Academic literature on the topic 'Humane Animal Treatment'

Animal mistreatment in businesses around the world is becoming a hotly debated topic. Many animal welfare laws protect wild animals and pets, but make exemptions for animals in farms, zoos or labs. There are economic benefits behind animal mistreatment since businesses can maximize profits by, for example, raising animals in crowded spaces, or forcing them to perform shows. However, ethical arguments on this issue reveal that animal mistreatment may actually cost more than humane animal treatment. Furthermore, consumer awareness on animal mistreatments is increasing, so this poses both a threat and opportunity to businesses. As society puts more and more value on sustainable green business today, inhumane animal treatments may harm a company's reputation and reduce its sales. Businesses should be aware of this trend and examine new humane alternatives to their traditional practices in order to stay competitive in the market.

AbstractResearch (Baldry, 2003; Flynn, 1999, 2000a; Henry, 2004) has linked witnessing abuse to nonhuman animals with the committal of such acts. This study reports frequency data based on adolescents' self-reported witnessing of animal abuse and involvement in animal-directed behaviors. The study investigates associations between witnessing abuse and engaging in both positive and negative animal-directed behaviors. 281 adolescents, 12-18 years of age, completed measures of animal cruelty and the humane treatment of animals. As predicted, the study found a history of witnessing animal abuse associated with significantly higher levels of animal cruelty. The study reported significantly higher levels of cruelty for those who had witnessed a friend, relative, parent, or sibling abuse an animal and significantly lower levels for those who had witnessed a stranger abuse an animal. Participants who "Frequently" witnessed animal abuse reported significantly higher levels of cruelty than those who viewed abuse "A few times". There was no association found between humane treatment of animals and the witnessing of animal abuse. Positive influences, peer mentors and humane education, would help to combat this cycle of abuse.

This essay examines several of the animal narratives that Margaret Marshall Saunders (1861–1947) wrote for children. From her first book, Beautiful Joe, a Dog’s Own Story, the first Canadian book to sell more than a million copies, to the end of her long career, Saunders developed a trenchant set of pedagogies and rhetorical strategies in support of humane education. Even as she negotiated her own culturally endorsed tendencies to see the animal as a thing, Saunders offered carefully reasoned arguments for the ethical treatment of animals through appealing pedagogies of humane education, renovations of her society’s views of non-human animals as objects for human consumption and pleasure, and rhetorical emphases on cultural understandings of connections between the child and the animal.

Animal Rights and Welfare: A Documentary and Reference Guide is a collection of fifty-one primary source documents relating to the topics of animal rights and animal welfare. The preface states that these are separate and distinct philosophies: animal rights advocates such as People for the Ethical Treatment of Animals and the Animal Liberation Front hold that humans and animals have the same rights (thereby precluding their use even as pets or assistive animals), whereas animal welfare adherents like the American Society for the Prevention of Cruelty to Animals and the American Humane Society endorse the use of animals for agriculture, work, biomedical research, etc., but in a manner that minimizes pain and suffering.

The article discusses some problems of legal regulation of the protection of farm animals, argues for the need to finalize the current legislation in terms of fixing legal norms in terms of humane treatment of this category of animals, justifies the need to take into account the best international experience in the field of humane treatment of animals. There is currently no single legislative act in the Russian Federation that would protect and support the rights of all animal species, including agricultural ones, to live satisfying their needs and experiencing minimal suffering. Recently, the changes taking place in the legislative regulation of relations in the field of humane treatment of animals have become more noticeable, which is expressed in the reform of this sphere of relations and the desire to bring it closer to the best world experience. However, to date, in Russia, no regulatory legal act has established rules that ensure humane conditions for keeping and slaughtering farm animals. The Federal Law "On Responsible Treatment of Animals" adopted in 2018 excluded farm animals from its scope, therefore, gaps in the legislative regulation.

The article discusses some problems of legal regulation of the protection of farm animals, argues for the need to finalize the current legislation in terms of fixing legal norms in terms of humane treatment of this category of animals, justifies the need to take into account the best international experience in the field of humane treatment of animals. There is currently no single legislative act in the Russian Federation that would protect and support the rights of all animal species, including agricultural ones, to live satisfying their needs and experiencing minimal suffering. Recently, the changes taking place in the legislative regulation of relations in the field of humane treatment of animals have become more noticeable, which is expressed in the reform of this sphere of relations and the desire to bring it closer to the best world experience. However, to date, in Russia, no regulatory legal act has established rules that ensure humane conditions for keeping and slaughtering farm animals. The Federal Law "On Responsible Treatment of Animals" adopted in 2018 excluded farm animals from its scope, therefore, gaps in the legislative regulation.

People can become infected with zoonoses from domestic or stray dogs (the most dangerous ones are rabies, brucellosis, dermatomycosis, leptospirosis, parasitic diseases). 1,466 rabies outbreaks and 2 cases of human rabies were reported in Ukraine in 2018. In total, 17 people died of rabies in Ukraine in 2014-2018. Sources of rabies were nonvaccinated against rabies dogs. In most dermatomycosis patients, the zoonotic form of microsporia caused by Microsporum canis is recorded, and infected cats and dogs play an important role in the spread. Dogs are a dangerous reservoir of leptospira and a source of infection for humans. Develops a mild course of leptospirosis, reminiscent of influenza, but has severe consequences and leads to meningitis. Dogs can infect humans with the agents of campylobacteriosis and brucellosis, as well as many other parasitic diseases: Ancylostoma braziliense і Ancylostoma caninum, Toxocara canis і Toxocara cati , Cryptosporidium , Echinococcus granulosus, Dirofilaria immitis , Giardia lamblia and ectoparasites: Sarcoptes scabiei (variant canis) and Cheyletiella yasguri. Prevention of zoonotic diseases requires a holistic approach within the framework of the One Health concept. Health, environmental and veterinary services, with the active involvement of local government, conservation and animal welfare organizations, must be involved. The problem of handling animals and regulating the numbers of stray animals in Sumy is an urgent one. The solution to this problem is to reduce the number of stray animals with only humane methods, to improve the epizootic and sanitary-ecological situation in the city, to create conditions for the prevention of the phenomenon of stray animals, and to change public opinion towards a civilized, humane, ethical attitude to animals. Principles of animal shelter organization, scheme of veterinary and sanitary measures and treatment and preventive treatment of animals have been developed and put into practice by us. Within the framework of the agreement with the communal enterprise of the Sumy City Council "Animal Care Center" Sumy National Agrarian University performs the veterinary part of the service of stray animals, which ensures the implementation of the "Program of regulation of the number of stray animals in the city of Sumy by humane methods".

The issue of how we ought to treat the nonhuman animals in our lives is one that has been growing in importance over the past forty years. A common charge is that discriminatory behavior based only on differences of species membership is just as wrong morally as are acts of racism or sexism. Is such a charge sustainable? It is argued that such reasoning confuses real differences with false ones, may have negative ethical consequences, and could tempt us to abandon our responsibilities to the natural world. Finally, some benefits to human animal treatment that more humane nonhuman animal treatment may bring are considered.

Alternative food systems (namely the humane product movement) have arisen to address societal concerns with the treatment of Nonhuman Animals in food production. This paper presents an abolitionist Nonhuman Animal rights approach (Francione, 1996) and critiques these alternative systems as problematic in regards to goals of considering the rights or welfare of Nonhuman Animals. It is proposed that the trend in social movement professionalization within the structure of a non-profit industrial complex will ultimately favor compromises like “humane” products over more radical abolitionist solutions to the detriment of Nonhuman Animals. This paper also discusses potential compromises for alternative food systems that acknowledge equal consideration for Nonhuman Animals, focusing on grassroots veganism as a necessary component for consistency and effectiveness.

This thesis has made a significant contribution to future chlamydial research by uncovering the chlamydial pathogenic mechanisms which will potentially help in the development of targeted vaccine against the pathogen. This thesis has made important new contributions to our understanding of Chlamydia pneumoniae specific adaptations to stress responses and has provided new perspectives on the survival of this successful pathogen. This thesis has used two well established microbial stressors and has identified major differences in stress responses between human and animal Chlamydia pneumoniae isolates.

Thesis (MSc (Physiological Sciences))--University of Stellenbosch, 2009.
Breast cancer is currently the primary cause of cancer-related death in women worldwide. Conventional treatments such as radiation and chemotherapy have many deleterious and long lasting side-effects, some of which are permanent, such as infertility. As certain tumour cells can also acquire resistance to chemotherapy, the need for the development of a less severe, yet more effective, targeted anti-cancer treatment exists. Curcumin, a plant polyphenol from Curcuma longa, has long been thought to possess antitumour, antioxidant, anti-arthritic, anti-amyloid, anti-ischemic and anti-inflammatory properties. Numerous studies conducted over the past sixty years confirm this. We aimed at examining the effect of curcumin on cell viability and the different modes of cell death, namely apoptosis, necrosis and autophagy, in the MCF-12A (non-tumorigenic mammary epithelial) and MCF-7 (mammary adenocarcinoma) cell lines. Cells were incubated with different doses of curcumin to evaluate the dose response through a MTT assay. Thereafter, cells were incubated with 200 μM curcumin for 48 hrs and stained with markers and DNA stains for apoptosis (Hoechst, Caspase-3, PARP), necrosis (Propidium Iodide) and autophagy (LC3B and Beclin-1). Cells were examined via fluorescence microscopy, Western Blot- and FACS analyses. MTT results showed no significant decrease in viability in the MCF-12A cell line after curcumin treatment. However, a significant decrease in viability was observed in MCF-7 cells after treatment with 200 μM curcumin (p < 0.05). Treated MCF-7 cells also show clear LC3B expression. FACS results show a significant difference in Hoechst mean fluorescence intensity in MCF-7 cells after curcumin treatment (p < 0.05). This study provides evidence that MCF-7 cells respond to a 200 μM dose of curcumin treatment through metabolic change and induction of the autophagic pathway. The model system used in this study provides groundwork for further cell culture based studies regarding breast cancer and curcumin.

It is difficult to overestimate the social and psychological significance of human-animal interactions. Till now, studies on human-companion animal interactions primarily focussed on positive aspects and relationships, while studies on animal phobias have almost exclusively focussed on spider and snake phobia. The problem with negative human-animal relationships in general, and animal phobia in particular, is in essence a superficial understanding of the determination of physiological changes and parameters associated with its description and treatment. The main aim of this study was to provide theoretical and physiological information regarding the determination of a biochemical parameter which can be used to enhance effective diagnosis and treatment of individuals suffering from dog phobia. A trimodal approach was followed to describe anxiety and fear responses associated with dog phobia. Subjects were assigned to two groups: an experimental group consisting of females suffering from dog phobia, and a control group. The study consisted of three experimental stages: the first stage (resting stage) measured baseline values, the second stage (preintervention stage) measured values in the presence of a dog stimulus prior to the intervention program, and the third stage (postintervention stage) measured values in the presence of a dog stimulus after completion of the intervention program. Cognitive-affective aspects were initially measured by means of the Fear Survey Schedule, as well as by means of an anxiety scale and stressor schedule during the experimental stages. Motor-behavioural aspects were measured as the termination distance of the dog approach during the pre- and postintervention stages, as well as assessed by a psychologist through direct observation of non-verbal communication cues during the behavioural approach tests. The measurement of physiological aspects focussed on the determination of plasma adrenocorticotropic hormone (ACTH) levels during the experimental stages. The main results were as follow: • the experimental group scored significantly higher average scores on the animal, dog, blood/injection and total fear categories of the Fear Survey Schedule than the control group; • the intervention program was effective in treating motor-behavioural and cognitive¬affective aspects of the phobia response; • the effect of the intervention program on the plasma ACTH-Ievels was inconclusive. No significant differences were found between the experimental group's average plasma ACTH-Ievels during the experimental stages, as well as between the experimental and control groups during the resting and preintervention stages. The average plasma ACTH-Ievels of the control group was significantly lower than that of the experimental group during the postintervention stage; • total stressor schedule values suggest that subjects in the experimental group have a predisposition to be generally more anxious and fearful than subjects in the control group; • two-thirds of the dog phobia subjects reported classical conditioning as the etiological pathway; • various auditory and visual cues were found to be the focal point of perception in women suffering from dog phobia; and • group qualitatively evaluated their current fear level for dogs as substantially lower than at the onset of the project. In conclusion, the determination of plasma ACTH-Ievels as a single parameter is not adequate to support the complex interaction between overt motor-behavioural, cognitive-affective and physiological patterns during the treatment of women experiencing dog phobia.
Dissertation (MSc (Veterinary Ethology))--University of Pretoria, 1999.
Production Animal Studies
unrestricted

Increasing evidence illustrates an involvement of stress in a large variety of physical and mental illness. Together with the evolutionary development of the social behavior in humans, the traditional interpretations of the attachment theory and the social support theory underscores the importance of affection, belonging and appreciation for human well-being. Not only can an imbalanced stress system be the cause of severe pathological consequences, insufficient social contact can also hamper recovery. Frequent usage of animals in various settings steadily illustrates both physiological and psychological benefits on both the young and the old, the healthy and the ill. Through the study of neurobiological factors, with oxytocin as a central mediator of social behavior and its  impact in turn on the stress- and cortisol system, this paper examines the possibility of animals to function as social support. The potential of animals to reduce the suffering in patients with stress related psychiatric disorders, such as the highly frequent exhaustion disorder, human-animal interactions might offer a non-invasive complementary tool to current treatment methods.

Thesis (PhD)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: Introduction Cisplatin has been widely used to treat solid tumours and much success has come from the use of this drug in the treatment of head and neck, ovarian, testicular, cervical and small-cell lung cancers. However, the success of cisplatin treatment is limited due to its dose-limiting toxicity and its resulting side-effects, such as nephro- and ototoxicity. The devastating side-effects induced by cisplatin treatment provided the platform for this study whereby the aim was to lower the concentration of cisplatin while maintaining its cancer-specific cytotoxic action. Equally concerning is, cisplatin resistance which is becoming increasingly common, and this radically limits the clinical efficacy and utility of the drug. Adjuvant therapy has thus become necessary in an attempt to possibly curb or lessen the extent of cisplatin resistance. Due to the large body of evidence implicating the importance of autophagy in cancer, the prospect of targeting this mechanism has generally been accepted. Various chemotherapy agents induce autophagy in cancer cells; however the effect of cisplatin on autophagic induction has not been very well explored. We thus hypothesise that the manipulation of the autophagic pathway will sensitise cancer cells to a low concentration of cisplatin treatment. Furthermore, due to the functional interaction between Bcl-2 and Beclin-1 and its role in the regulation of autophagy, ratio analysis of Beclin-1 to Bcl-2 as means of detecting the role of autophagy within the cell under homeostatic and treatment/stress conditions has been conducted. Additionally, Bcl-2 has a prominent role in the malignant cell and it’s over-expression has been found to confer resistance in a variety of cancerous cell lines. We therefore hypothesise that the silencing of Bcl-2 prior to cisplatin treatment will sensitise cervical cancer cells to apoptosis and increase the Beclin-1/Bcl-2 ratio in favour of apoptosis. Materials and Methods Three human cervical cell lines were used: a non-cancerous ectocervical epithelial cell line (Ect1/E6E7) and two cancerous cervical cell lines (HeLa and CaSki). In order to determine a concentration of cisplatin that was non-toxic to the non-cancerous Ect1/E6E7 cell line, a dose-response was performed. With the use of an autophagy inhibitor (bafilomycin A1) and an autophagy inducer (rapamycin), autophagic flux capacities were assessed in each cell line through the Western blotting technique. In order to assess whether the chosen concentration of cisplatin induced autophagy, flow cytometry with the use of a Lysotracker™ dye was utilised, as well as analysis of autophagy protein levels (LC-3 II, Beclin-1 and p62). Autophagy modulation was achieved through two methods: pharmacological modulation with use of two recognised agents, namely bafilomycin A1 and rapamycin, and biological manipulation with the use of ATG5 and mTOR mRNA silencing. The effects of different treatment regimes on cell death was assessed with the use of PARP and caspase-3 cleavage through Western blotting, caspase-3/-7 activity (Caspase-Glo®), PI inclusion, LDH release and MTT reductive capacity. Additionally the effects of these treatment regimes on cell-cycle progression were also analysed. Beclin-1 and Bcl-2 expression was determined through Western blotting and immunocytochemistry before and after treatment with cisplatin in HeLa and CaSki cells. To assess the reliance of the cervical cancer cells on Bcl-2 after cisplatin treatment, Bcl-2 knock-down was achieved through RNA interference, where after the Beclin-1/Bcl-2 ratio was assessed as well as apoptosis with the use of cleaved PARP analysis (Western blotting) and Caspase-Glo©. For the ex vivo analysis, biopsies were collected from patients undergoing routine colposcopy screenings and hysterectomies at Tygerberg Hospital, Tygerberg, Western Cape. A total of 10 normal, 29 low-grade squamous intraepithelial lesions (LSIL), 33 high-grade squamous intraepithelial lesions (HSIL) and 13 carcinoma biopsies were collected for analysis, where after the expression profiles of two autophagy markers (mTOR and LC-3 II), as well as one anti-apoptotic marker (Bcl-2) were assessed. Protein levels were analysed through Western blot and confirmed through immunohistochemistry. Results Dose-response curves revealed that 15 μM of cisplatin did not induce cell death in the normal cervical epithelial cell line (Ect1/E6E7) and was therefore utilised through-out the remainder of the study. It was additionally determined that the CaSki cells were more resistant to cisplatin treatment when compared to the HeLa and Ect1/E6E7 cells. Autophagic flux analysis revealed that, although all three cell lines were cervix derived, their autophagic flux capacities differed. It was observed that the chosen concentration of cisplatin was able to induce autophagy in all three cell lines, with the HeLa cells demonstrating a particularly pronounced response. Autophagy modulation in conjunction with cisplatin treatment revealed the following: Autophagy inhibition with bafilomycin A1 lead to significant increases in caspase-3 and PARP cleavage and LDH release in both cervical cancer cell lines. The inhibition of autophagy through silencing of ATG5 induced caspase-3 cleavage and agrees with results obtained from pharmacological inhibition of autophagy with bafilomycin A1. In addition to autophagic induction, a low concentration of cisplatin induced the up-regulation of Bcl-2, which when silenced significantly improved cisplatin-induced apoptosis in both cervical cancer cell lines. Analysis of the expression profiles of mTOR and LC-3 in normal, pre-malignant (LSIL and HSIL) and cancerous cervical tissue revealed that autophagy is significantly up-regulated in HSILs and carcinoma of the cervix. Additionally, Bcl-2 expression is significantly increased in cervical carcinoma tissue, which agrees with results from other studies. Conclusion Autophagic flux capacities between the three cell lines investigated, derived from the same organ, differ significantly. This should be taken into consideration when autophagic modulation is being used as an adjuvant treatment. With regard to chemotherapy treatment in cervical cells, a low-concentration of cisplatin significantly induces autophagy in malignant and non-malignant cervix-derived cell lines where it serves a pro-survival mechanism. Inhibition of autophagy with bafilomycin A1 and ATG5 siRNA confirmed this survival effect in both cancerous cell lines where apoptosis was significantly increased. Interestingly, rapamycin pre-treatment together with cisplatin did not induce significant levels of apoptosis in HeLa cells where autophagy induction may have provided additional protection from the cytotoxic effects of cisplatin. Therefore the inhibition of autophagy through pharmacological and biological inhibition improves the cytotoxicity of a low concentration of cisplatin and provides a promising new avenue for the future treatment of cervical cancer. Bcl-2 up-regulation in response to cisplatin treatment also serves as a protective mechanism by which cervical cancer cells survive. The extent of apoptotic cell death observed after biological inhibition of Bcl-2 reiterates the fact that this response may be exploited in order to favour the use of lower concentrations of cisplatin. Analysis of clinical specimens emphasised the value of the in vitro work: Cervical cancer biopsies had increased expression of both LC-3 II and Bcl-2, indicating autophagy induction and apoptosis inhibition, respectively. Thus two novel methods of improving cisplatin cytotoxicity have been demonstrated in the following study. Treatment regimens may administer more frequently and prolonged due to the minimal side-effects that accompanies low-dose cisplatin treatment.
AFRIKAANSE OPSOMMING: Inleiding Sisplatien word algemeen gebruik vir die behandeling van soliede gewasse. Baie sukses is reeds deur die gebruik van díe middel behaal in die behandeling van kop en nek, ovariale, terstikulêre, servikale en klein-sel kankers. Die sukses van Sisplatien-behandeling word wel ingeperk deur die dosis-beperkende toksisiteit en die gevolglike newe-effekte soos nefrotoksisiteit. Hierdie verwoestende newe-effekte wat deur sisplatien behandelings geïnduseer word, het as die platform vir hierdie studie gedien. Die doel was om die sisplatien konsentrasies te verlaag, maar terselfdertyd die kankerspesifieke sitotoksisiteit te behou. Nog ʼn punt van kommer is dat sisplatien-weerstandigheid aan die toeneem is, wat die kliniese effektiwiteit en gebruik van hierdie middel geweldig beperk. Byvoegmiddels het dus noodsaaklik geraak in die poging om die sisplatien-weerstandigheid te verhoed. As gevolg van verskeie bewyse wat die belangrikheid van outofagie in kanker impliseer, is die vooruitsig om hierdie meganisme te teiken, algemeen aanvaar. Verskeie chemoterapeutiese middels induseer outofagie in kanker selle, hoewel die effek van Sisplatien op outofagiese induksie nog nie goed ondersoek is nie. Ons hipotese is dus dat die manipulasie van die outofagiese pad die kankerselle sensitiseer tot ʼn lae konsentrasie van sisplatien. Verder, as gevolg van die funksionele interaksie tussen Bcl-2 en Beclin-1, en hul rol in die regulering van outofagie, is verhouding-analises van Beclin-1 tot Bcl-2 uitgevoer met die doel om die rol van outofagie in die sel onder homeostatiese en behandeling/stres kondisies te bepaal. Verder is Bcl-2 bekend daarvoor om ʼn prominente rol te speel in kwaadaardige selle, en die ooruitdrukking daarvan is gevind om weerstandigheid aan te help in ʼn verskeidenheid van kankeragtige sellyne. Ons hipotetiseer dus dat geenonderdrukking van Bcl-2 voor die behandeling met sisplatien die servikale kanker selle sal sensitiseer tot apoptose en ʼn verhoging in die verhouding van Beclin-1/Bcl-2 veroorsaak, wat in die guns van apoptose is. Materiale en Metodes Drie menslike servikale sellyne was gebruik: ʼn nie-kankeragtige servikale epiteel sellyn (Ect/E6E7) en twee kankeragtige servikale sellyne (HeLa en CaSki). Om ʼn konsentrasie van sisplatien te bepaal wat nie-toksies tot die nie-kankeragtige Ect1/E6E7 sellyn is, was ʼn dosisrespons uitgevoer. Met die gebruik van ʼn outofagiese inhibeerder (bafilomycin A1) en ʼn outofagiese induseerder (rapamycin), is die outofagiese-fluks kapasiteite van elke sellyn deur die Western Blotting tegniek geassesseer. Om te bepaal of die gekose konsentrasie van sisplatien outofagie induseer, is vloeisitometrie met ʼn Lysotracker™ kleurstof gebruik, sowel as analises op outofagie proteïenvlakke (LC-3 II, Beclin-1 en p62). Outofagie modulering is behaal deur twee metodes: farmakologiese modulering met twee erkende middels, naamlik bafilomycin A1 en rapamycin, en biologiese manipulasie met die gebruik van ATG5 en mTOR geenonderdrukking. Die effekte van die verskillende behandeling skedules op seldood was geassesseer deur gebruik te maak van PARP en kaspase-3 splitsing deur Western Blotting, kaspase-3/-7 aktiwiteit deur Caspase-Glo ®, PI-insluiting, LDH vrystelling en MTT reduserende kapasiteit. Verder is die effekte van hierdie behandeling skedules op selsiklus progressie ook geanaliseer. Beclin-1 en Bcl-2 uitdrukking was ook bepaal deur Western Blotting en immunohistochemie voor en na behandeling met sisplatien in HeLa en CaSki selle. Om die afhanklikheid van die servikale kankerselle op Bcl-2 na sisplatien behandelings te toets, is Bcl-2 onderdruk deur RNA-inmenging, waarna Beclin-1/Bcl-2 verhouding geassesseer is, sowel as opoptose deur die gebruik van gesplitste PARP analises (Western Blotting) en Caspase-Glo©. Vir die ex vivo analises is biopsies vanaf pasiënte wat roetine kolposkopie en histerektomies ondergaan, verkry (Tygerberg Hospitaal, Tygerberg, Westelike Provinsie). ʼn Totaal van 10 normale, 29 lae-graad plaveisel intraepiteel letsels (LSIL), 33 hoe-graad plaveisel intraepiteel letsels (HSIL) en 13 karsinoom biopsies is verkry vir analises. Die uitdrukkingsprofiel van twee outofagiese merkers (mTOR en LC-3 II), asook een merker vir apoptose (Bcl-2), was geassesseer. Proteïen vlakke was ook deur Western Blotting geanaliseer en deur immunohistochemie bevestig. Resultate Dosisrespons kurwes het getoon dat 15 μM sisplatien nie seldood in die normale sellyn (Ect1/E6E7) geïnduseer het nie, en was daarom gebruik deur die res van hierdie studie. Verder is daar ook gevind dat CaSki selle meer weerstandig tot sisplatien behandelings is wanneer vergelyk word met die HeLa en Ect1/E6E7 selle. Outofagiese-fluks analises het getoon dat, alhoewel al drie sellyne vanaf die serviks afkomstig is, daar verskille is in hul outofagiese-fluks kapasiteit. Daar is ook waargeneem dat die gekose konsentrasie van sisplatien in staat was om outofagie te induseer in al drie sellyne, met HeLa selle wat die mees merkbare respons getoon het. Modulering van outofagie in samewerking met sisplatien behandelings het die volgende onthul: inhibisie van outofagie deur bafilomycin A1 het gelei tot ʼn beduidende verhoging in kaspase-3, PARP splitsing en LDH vrylating in beide servikale kankersellyne. Geenonderdrukking van ATG5 induseer kaspase-3 splitsing en stem ooreen met resultate wat verkry is deur farmakologiese inhibisie van outofagie met bafilomycin A1. Bykomend tot outofagiese indusering, het ʼn lae konsentrasie sisplatien die opregulering van Bcl-2 geïnduseer. Wanneer Bcl-2 geenonderdrukking in hierdie scenario toegepas was, het dit ʼn beduidende verbetering in sisplatien-geïnduseerde apoptose in beide servikale kankersellyne getoon. Analises van die uitdrukkingsprofiel van mTOR en LC-3 in normale, pre-maligne (LSIL en HSIL) en kankeragtige servikale weefsel, het getoon dat outofagie beduidend opgereguleer is in HSILs en servikale karsinome. Verder is Bcl-2 uitdrukking ook gevind om beduidend verhoog te wees in servikale karsinoomweefsel, wat ooreenstem met resultate verkry in ander studies. Gevolgtrekking Outofagiese-fluks kapasiteite tussen die drie sellyne, afkomstig van dieselfde orgaan, toon beduidende verskille. Hierdie bevinding moet in ag geneem word wanneer outofagiese-modulering as ʼn bevorderingsbehandeling gebruik word. Met betrekking tot chemoterapie behandeling in servikale selle; ʼn lae konsentrasie van sisplatien veroorsaak ʼn beduidende indusering van outofagie in kwaadaardige en nie-kwaadaardige serviks-afkomstige sellyne, en dien as ʼn oorlewingsmeganisme. Inhibisie van outofagie met bafilomycin A1 en ATG5 siRNA het hierdie beskermings effek bevestig, aangesien apoptose beduidend verhoog was in beide kankersellyne. Interessant genoeg het rapamycin pre-behandeling tesame met sisplatien nie beduidende vlakke van apoptose in HeLa selle geïnduseer nie. Outofagie induksie mag dalk addisionele beskerming teen die sitotoksiese effekte van sisplatien gebied het. Daarom het die inhibisie van outofagie deur farmakologiese en biologiese inhibering die sitotoksisiteit van ʼn lae konsentrasie sisplatien bevorder, wat ʼn belowende bevinding is vir die toekomstige behandeling van servikale kanker. Bcl-2 opregulering as gevolg van sisplatien behandelings dien ook as beskermings meganisme waarby servikale kankerselle oorleef. Die mate van apoptotiese seldood wat waargeneem word na biologiese inhibering van Bcl-2, wys weer op die feit dat hierdie respons uitgebuit kan word vir die gebruik van laer konsentrasies van sisplatien. Analises van die kliniese monsters het ook die waarde van die in vitro werk versterk: Servikale kanker biopsies het verhoogde uitdrukking van beide LC-3 II en Bcl-2 getoon, wat aandui dat outofagie geïnduseer en apoptose geïnhibeer word. Daar is dus twee nuwe metodes vir die verbetering van sisplatien-toksisiteit in hierdie studie gedemonstreer. Behandeling regimes kan meer gereeld en vir langer tydperke toegepas word, aangesien die newe-effekte van lae-dosis sisplatien behandelings minimaal is.
MRC for funding

This study aims to deconstruct current conceptions about animal-assisted interventions by investigating relationships between human beings and birds of prey. Interactions between birds of prey, or “raptors,” provide novel cases from which to reexamine failed attempts to provide empirical data in support of alternative therapies. Previous research addressing the efficacy of animal-assisted interventions is simply not robust enough to be considered a feasible treatment option by medical professionals. By extension, models of self-regulation in psychology are often presented using reductionist models and oversimplified therapeutic outcomes. Taken together, raptor-human relationships help to highlight the shortcomings of each, as well as potential solutions towards developing comprehensive frameworks for measuring efficacy of multispecies interactions. This study was conducted at a small nature park in Largo, FL where a number of native raptor species are housed, cared for, and trained each day by volunteers. These volunteers made up the sample size for this study with forty participants (n = 40) between the ages of eighteen and seventy five. Drawing on both my own experiences as a raptor handler, as well as the qualitative data collected from volunteers, I employed a neuroanthropological approach to reveal underlying dynamics of the program via a two-stage research plan. Stage 1 of the study addresses the Raptor Program itself in facilitating human-animal interactions. Stage 2 addresses the mechanisms at play during firsthand encounters with birds of prey. Findings suggest that programmatic and regulatory drivers within the program must operate together, often simultaneously, for an animal-assisted intervention organization to be successful. Further, this study calls for the ongoing development of novel methodological approaches in future research to determine the efficacy of animal-assisted interventions at large.

AbstractAnimals, like human beings, are prone to suffering harms, such as disease, injury and death, as a result of anthropogenic and natural disasters. Animals are disproportionately prone to risk and adversely affected by disasters, and thus require humane and respectful care when disasters strike, due to socially situated vulnerabilities based on how human communities assess and value their moral standing and function. The inability to integrate animals into disaster risk and management practices and processes can sometimes be associated with a lack of understanding about what animal ethics and animal health and welfare require when designing disaster management programs. This chapter seeks to reimagine human responsibility towards animals for disaster management. The pervasiveness of disasters and their impacts on animals, human-animal and animal-environment relationships underscore the importance of effective animal disaster management supported by sound ethical decision-making processes. To this end, we delineate six ethically responsible animal caretaking aims for consideration when developing disaster management plans and policies. These aims, which address central vulnerabilities experienced by domesticated animals during disasters, are meant to be action-guiding within the disaster management context. They include: (1) Save lives and mitigate harm; (2) Protect animal welfare and respect animals’ experiences; (3) Observe, recognize and promote distributive justice; (4) Advance public involvement; (5) Empower caregivers, guardians, owners and community members; (6) Bolster public health and veterinary community professionalism, including engagement in multidisciplinary teams and applied scientific developments. To bring about these aims, we offer a set of practical and straightforward action steps for animal caregivers and disaster management teams to ensure that animals’ interests are systematically promoted in disaster management. They include: (1) Respect and humane treatment; (2) Collaboration and effective disaster communication; (3) Strengthening systems of information sharing, surveillance, scientific research, management and training; (4) Community outreach and proactive contact; (5) Cultural sensitivity and attitudes check, and (6) Reflection, review and reform.

AbstractWithin the animal movement there is a tension between those who favor a ‘humane treatment’ of animals and those who think we need to abolish the (ab)use of animals all together and grant animals legal personhood. For an number of reasons philophers and other reseachers often try not to take a stance in this strategy debate. They however fail in doing so. As they make recommendations on how to treat animals they cannot avoid taking an implicit of explicit stance in the debate. This paper argues that we need a ‘directionists approach’ in this matter. We, as philosophers or reseachers, may for all sorts of reasons advice farmers, organisations or governments to take small steps towards more animal welfare, but only if we make clear on the outset that this will not be the end of the journey. We, as a society and as philosophers and researchers, are heading in a direction and one can either take the long and exhausting road of incremental change or take a shortcut.

Abstract This chapter first outlines a trend of change in the image of animals in several disciplines, which indicates the emergence of a paradigmatic shift in the treatment of animals and their relations to humans. It then discusses the apparent deficiencies in the study of tourist-animal relations, and ends up with some suggestions to rethink the basic assumptions and theoretical approaches to animals in tourism studies.

Abstract People may have preferences in relation to products and services that they perceive to have animal welfare attributes. People may also have preferences in relation to different welfare states, standards and treatment of animals. Human empathy with animals is a potential source of utility (e.g. the pleasure from seeing animals playing) and disutility (e.g. the distress caused if we perceive an animal is suffering). We may derive satisfaction from moral preferences for how animals are treated regardless of this empathy. There are many animal-derived products for consumers to purchase in markets but little information on the welfare of the animals that produced them in order for consumers to make choices consistent with their animal welfare preferences. People's preferences can be ascertained by observations of their behaviour and the choices they make, and by asking people what their preferences are. Two research case studies are presented. There is a need for welfare advocacy on behalf of animals. This stewardship role must be informed by scientific evidence on animal sentience, preferences and welfare status. Government or an alternative body acting on behalf of society must take up the role to represent societal animal welfare preferences and act as custodians to protect the welfare of animals.

Abstract Glioblastoma Multiforme (GBM) is a malignant brain cancer with low overall survival. Therefore, researchers are looking to augment its current therapeutic regimen, which includes surgical tumor resection, chemotherapy and radiation. A promising treatment modality, focused ultrasound, has been used as a non-invasive treatment for GBM through multiple approaches such as thermal ablation, immunomodulation, and blood brain barrier disruption. In order to develop these treatments for clinical trials, testing in animal models needs to be performed to investigate the efficacy of the treatment in complex biological environments, as well as to evaluate any side-effects. The more biologically relevant the animal model is to human anatomy, the more applicable the results will be for translation to clinical trials. Here, we report a human GBM rat model, which utilizes an IDH-wildtype, EGFRvIII mutant patient-derived xenograft in athymic rats. The in vivo tumor growth rate was assessed over a period of 20 days to evaluate reproducibility and to develop the model for future testing of FUS in the treatment of GBM.

Org 10172 is a novel low molecular weight heparinoid isolated from animal mucosa with a higher anti-thrombotic/bleeding risk ratio than standard heparin. The effects of Org 10172 and heparin on primary haemostasis in rats and humans have been studied by light and electron microscopy (LM&EM).In rat ears bleeding wounds were inflicted. The wound areas were excised 5, 15 or 30 min. after bleeding induction, and processed for LM and EM. Org 10172 and heparin have been administered in doses of 300 or 600 anti-Xa U/kg i.v., 5 or 1 min. prior to bleeding induction. Bleeding wounds of drug treated animals have been compared with those of placebo animals. Heparin inhibited degranulation as well as fibrin deposition and groups of not aggregated, sometimes even discoidal platelets could be found after heparin treatment. Org 10172 inhibited degranulation to some extent but less than heparin and Org 10172 hardly inhibited fibrin deposition.In six human volunteers SimplateR bleeding time wounds were excised by punch biopsy, 20 min. after bleeding induction and processed for LM and EM. Org 10172 was administered as single bolus injection of either 3200 or 6400 anti-Xa U i.v., 10 min. prior to bleeding induction. Heparin was given in a dosis of 10.000 anti-Xa U i.v. All biopsies from post-drug bleeding time wounds were compared with biopsies taken from pre-drug bleeding time wounds in the same volunteer. As in the rat studies, heparin inhibited degranulation and fibrin deposition whereas after Org 10172 treatment these effects were hardly detectable. In general the effects of both drugs on haemostasis in the human volunteer study were less distinct than in the rat study. This may be attributed to the lower dose levels used.In conclusion, primary haemostasis after Org 10172 treatment is somewhat retarded, but essentially normal, whereas haemostasis after heparin treatment is more severely disturbed.

Newly developed cancer therapies must pass through a series of increasingly complex testing regimens before obtaining FDA approval as valid treatments. The costs of these tests increase rapidly as the physiological accuracy of the platform increases, from initial proof-of-concept in static tissue cultures, to treatment of animal models, and ultimately to human clinical trials. Three-dimensional engineered blood-perfused tumor models are becoming increasingly important as intermediate platforms for the study and treatment of cancer, as they are superior to static two-dimensional cultures in their reproduction of relevant physiological conditions and are inexpensive in comparison to animal models. Because of this, the design of well-characterized adaptable in vitro vascular tumor models has become a central objective of the emerging field of tumor engineering. Characterization of the flow within three-dimensional tumor models is critical for quantifying fluid shear stress and determining its role in pivotal tumor development processes such as tumor cell angiogenesis and metastasis. Ultimately, this knowledge will provide new avenues for therapeutic modulation of the tumor microenvironment.

Diagnosis, repair and regeneration of the disc often necessitate needle injection to the nucleus pulposus through the annulus. Discography in which a radio opaque material is injected into the nucleus and electrothermal treatment involving inserting a catheter into the disc requires disruption of the annulus through needle puncture. Annulus puncture may also be required during placement of nucleus implants. Needle puncture is also used to inject growth factors, gene and cell therapy for regeneration of the disc. In animal models, disc degeneration is induced over time by needle puncture of the annulus. The severity of the degeneration depends on the magnitude of the annulus needle puncture. One thing that is not clear is how much of the observed changes in the disc biomechanics and biochemical changes are due to nucleus treatment and how much is due to annular disruption through needle puncture. Animal model studies have shown that significant changes in disc mechanics were noticed within 1 week of needle puncture with a large-gauge needle. Another in-vitro animal study showed that biomechanical changes were observed in the disc when the ratio of needle diameter to disc height is greater than 40%. All these studies were focused on the effect of small number of needle diameters and addressed using animal cadaver models. How these needle puncture injury studies on small and large animal models can be extrapolated to human conditions is still not known. Thus there is need to evaluate effect of range of needle puncture diameters in human lumbar disc biomechanics. The purpose of this study is, with the help of a finite element models, quantify the biomechanical effect due to varying size of needle punctures in a human lumbar intervertebral disc.

Patients frequently experience knee injuries, with the ACL being one of the most commonly injured structures requiring surgery [1]. ACL tears typically lead to osteoarthritis in the long term, even after surgical treatment [2]. This chronic joint degeneration has been attributed to the failure of current ACL reconstructions to restore the native biomechanics of the knee joint [3]. To design more effective treatments, investigators must first understand normal knee function for multiple activities of daily living (ADLs). The 3D in vivo forces and moments of the normal intact knee, as well as those for just the ACL have not yet been determined for any ADL. These in vivo forces and moments can potentially be measured for multiple ADLs in an animal model. A biomechanical surrogate allows for 1) sensors or marker systems to be rigidly fixed to the knee joint to accurately measure the 6 degree of freedom (DOF) kinematics, and for 2) the kinematics to be simulated and applied to the harvested limb to measure the corresponding joint forces and moments.

Background: Parkinson’s disease (PD) is a neurological disorder that affects movement, mainly due to damage and degeneration of the nigrostriatal dopaminergic pathway. The diagnosis is made through a clinical neurological analysis where motor characteristics are considered. There is still no cure, and treatment strategies are focused on symptoms control. Cell replacement therapies emerge as an alternative. Objective: This review focused on current techniques of induced pluripotent stem cells (iPSCs). Methods: The search terms used were: “Parkinson’s Disease”, “Stem cells” and “iPSC”. Open articles written in English, from 2016-21 were selected in the Pubmed database, 10 publications were identified. Results: With the modernization of iPSC, it was possible to reprogram pluripotent human somatic cells and generate dopaminergic neurons and individual-specific glial cells. To understand the molecular basis, cell and animal models of neurons and organelles are currently being employed. Organoids are derived from stem cells in a three-dimensional matrix, such as matrigel or hydrogels derived from animals. The neuronal models are: α-synuclein (SNCA), leucine-rich repeat kinase2 (LRRK2), PARK2, putative kinase1 induced by phosphatase and tensin homolog (PINK1), DJ-1. Both models offer opportunities to investigate pathogenic mechanisms of PD and test compounds on human neurons. Conclusions: Cell replacement therapy is promising and has great capacity for the treatment of neurodegenerative diseases. Studies using iPSC neuron and PD organoid modeling is highly valuable in elucidating relevants neuronal pathways and therapeutic targets, moreover providing important models for testing future therapies.

Intensive studies were reported on the osteogenesis of mesenchymal stem cells (MSC) using chemicals and mechanical loading. However, the maturity of differentiated osteoblasts is not same as that of isolated adult osteoblasts. Thermal treatment could be a missing factor in stem cell differentiation. It was reported that mild heat stimulated bone growth in animal experiments [1–2]. Thermal treatment is also used as a therapy to promote bone repair after injury [3]. In addition, hot shower daily is recommended to osteoarthritis patients. However, the mechanisms for the heat-induced osteogenesis are not completely known and the thermal regulation of human mesenchymal stem cells (hMSCs) differentiation is not well studied. In this study, the direct effects of mild heat shock (HS) on the differentiation of hMSCs into osteoblasts in self-assembling peptide hydrogel were investigated.

Human infection with Trypanosoma parasites (Chagas disease and Human African Trypanosomiasis) affects around 10 million people worldwide resulting in life-threatening disease. Treatment options are limited to historic drugs characterized by significant side effects and decreasing efficacy while new drug development efforts are largely neglected. Here, we review drug discovery effort in human trypanosomiasis undertaken in academia. Peroxisomal (Pex) transport system was validated as a target in Chagas disease and a number of compounds were delivered which have shown promising results in animal experiments. Future perspectives of exploring the Pex system in anti-trypanosoma drug development are discussed.

Traditionally, lavender is believed to have various therapeutic and curative properties, ranging from inducing relaxation and ending with the treatment of parasitic infections, burns, insect bites and spasms. There is more and more evidence that lavender oil can be an effective drug in the treatment of certain neurological disorders. Some animal and human studies point to the anxiolytic, mood-stabilizing, sedative, analgesic, anticonvulsant and neuroprotective properties of lavender. These studies have raised the possibility of reviving the therapeutic efficacy of lavender in neurological disorders. This article presents an overview of the current experimental and clinical state of knowledge about the effect of lavender on the nervous system.

Drug development platforms such as two-dimensional (2D) in vitro cell culture systems and in vivo animal studies do not accurately predict human in vivo effectiveness of candidate therapeutics [1]. Cell culture systems have limited similarities to primary human cells and tissues as only one cell type is employed and animal studies have a generally limited ability to recapitulate human drug response as different species have differences in metabolism, physiology, and behavior. Mike Leavitt, a former U.S. Secretary of Health and Human Services, has stated that “currently, nine out of ten experimental drugs fail in clinical studies because we cannot accurately predict how they will behave in people based on laboratory and animal studies” [2]. Therefore, this research project is focused on developing an in vitro platform to test candidate therapeutics for more efficacious predictions of human response. We have fabricated a three-dimensional (3D) breast cancer tissue volume containing a vascular network. This vascular network is necessary because current in vitro systems (e.g., rotating bioreactors, suspension of spheroids, and growth on a porous scaffold) are limited in size (1–2 mm) by their absence of micrometer-scale blood flow micro-channels that allow for oxygen and nutrient diffusion into the tissue [4]. The extracellular matrix scaffold has been developed to mimic the native extracellular matrix and includes relevant cell types (e.g., human breast cancer epithelial cells and human breast fibroblasts) along with the prefabricated vascular network (prevascularization). These systems are intended to support long-term growth, recapitulate physiological tissue function, and accurately model response to treatment. It is hypothesized that the development of reproducible tissue volumes will transform breast cancer drug development by providing reliable, cost-effective models that can more accurately predict therapeutic efficacy than current preclinical in vivo and in vitro models.

The goal for this manual is to provide practical information for the implementation of mitigation measures that aim to: 1. Improve human safety through reducing collisions with large animals, including large wild mammal species, select free roaming large feral species, and select free roaming large livestock species, and 2. Improve or maintain habitat connectivity for terrestrial wildlife species and selected feral species through safe crossing opportunities. This manual does not include all possible measures that can or may reduce animal-vehicle collisions and maintain or improve habitat connectivity for wildlife. The measures included in this manual are: Barriers (fences) in combination with crossing structures (for large wild mammals and for small wild animal species), roadside animal detection system, Barriers (fences), Barriers (fences) in combination with crossing structures (for free roaming livestock), and culling, relocation, anti-fertility treatment, roadside animal detection systems, barriers (fences), and barriers (fences) in combination with crossing structures (for large feral mammal species such as feral horses and burros).

Addressing the public health threat posed by AMR is a national strategic priority for the UK, which has led to both a 20-year vision of AMR and a 5-year (2019 to 2024) AMR National Action Plan (NAP). The latter sets out actions to slow the development and spread of AMR with a focus on antimicrobials. The NAP used an integrated ‘One-Health’ approach which spanned people, animals, agriculture and the environment, and calls for activities to “identify and assess the sources, pathways, and exposure risks” of AMR. The FSA continues to contribute to delivery of the NAP in a number of ways, including through furthering our understanding of the role of the food chain and AMR.Thorough cooking of food kills vegetative bacterial cells including pathogens and is therefore a crucial step in reducing the risk of most forms of food poisoning. Currently, there is uncertainty around whether cooking food is sufficient to denature AMR genes and mobile genetic elements from these ‘dead’ bacteria to prevent uptake by ‘live’ bacteria in the human gut and other food environments - therefore potentially contributing to the overall transmission of AMR to humans. This work was carried out to assess these evidence gaps.

Objectives: (1) Evaluate Immunocompetence-OTL-containing Chromosomal Regions (ICRs), marked by microsatellites or candidate genes, for magnitude of direct effect and for contribution to relationships among multiple immunocompetence, disease-resistance, and growth traits, in order to estimate epistatic and pleiotropic effects and to predict the potential breeding applications of such markers. (2) Evaluate the interaction of the ICRs with genetic backgrounds from multiple sources and of multiple levels of genetic variation, in order to predict the general applicability of molecular genetic markers across widely varied populations. Background: Diseases cause substantial economic losses to animal producers. Emerging pathogens, vaccine failures and intense management systems increase the impact of diseases on animal production. Moreover, zoonotic pathogens are a threat to human food safety when microbiological contamination of animal products occurs. Consumers are increasingly concerned about drug residues and antibiotic- resistant pathogens derived from animal products. The project used contemporary scientific technologies to investigate the genetics of chicken resistance to infectious disease. Genetic enhancement of the innate resistance of chicken populations provides a sustainable and ecologically sound approach to reduce microbial loads in agricultural populations. In turn, animals will be produced more efficiently with less need for drug treatment and will pose less of a potential food-safety hazard. Major achievements, conclusions and implications:. The PI and co-PIs had developed a refined research plan, aiming at the original but more focused objectives, that could be well-accomplished with the reduced awarded support. The successful conduct of that research over the past four years has yielded substantial new information about the genes and genetic markers that are associated with response to two important poultry pathogens, Salmonella enteritidis (SE) and Escherichia coli (EC), about variation of immunocompetence genes in poultry, about relationships of traits of immune response and production, and about interaction of genes with environment and with other genes and genetic background. The current BARD work has generated a base of knowledge and expertise regarding the genetic variation underlying the traits of immunocompetence and disease resistance. In addition, unique genetic resource populations of chickens have been established in the course of the current project, and they are essential for continued projects. The US laboratory has made considerable progress in studies of the genetics of resistance to SE. Microsatellite-marked chromosomal regions and several specific genes were linked to SE vaccine response or bacterial burden and the important phenomenon of gene interaction was identified in this system. In total, these studies demonstrate the role of genetics in SE response, the utility of the existing resource population, and the expertise of the research group in conducting such experiments. The Israeli laboratories had showed that the lines developed by selection for high or low level of antibody (Ab) response to EC differ similarly in Ab response to several other viral and bacterial pathogens, indicating the existence of a genetic control of general capacity of Ab response in young broilers. It was also found that the 10w-Ab line has developed, possibly via compensatory "natural" selection, higher cellular immune response. At the DNA levels, markers supposedly linked to immune response were identified, as well as SNP in the MHC, a candidate gene responsible for genetic differences in immunocompetence of chickens.

Original Objectives. The overall goal is to develop methods to increase pregnancy rate in lactating dairy cows exposed to heat stress through methods that minimize damage to the oocyte and embryo caused by heat stress. Objectives were as follows: (1) examine the protective effects of melatonin on developmental competence of oocytes exposed to elevated temperature in vitro; (2) test whether melatonin feeding can improve developmental competence of oocytes in vivo and, if so, whether effects are limited to the summer or also occur in the absence of heat stress; and (3) evaluate the effectiveness of improving fertility by facilitating follicular turnover in the summer and winter. Revised Objectives. (1) Examine protective effects of melatonin and follicular fluid on developmental competence of oocytes exposed to elevated temperature in vitro; (2) examine the protective effects of melatonin on developmental competence of embryos exposed to elevated temperature in vitro; (3) evaluate effectiveness of improving fertility by administering human chorionicgonadotropin (hCG) to increase circulating concentrations of progesterone and evaluate whether response to hCG depends upon genotype for four mutations reported to be related to cow fertility; and (4) identify genes with allelic variants that increase resistance of embryos to heat shock. Background. The overall hypothesis is that pregnancy success is reduced by heat stress because of damage to the oocyte and cleavage-stage embryo mediated by reactive oxygen species (ROS), and that fertility can be improved by provision of antioxidants or by removing follicles containing oocytes damaged by heat stress. During the study, additional evidence from the literature indicated the potential importance of treatment with chorionicgonadotropin to increase fertility of heat- stressed cows and results from other studies in our laboratories implicated genotype as an important determinant of cow fertility. Thus, the project was expanded to evaluate hCG treatment and to identify whether fertility response to hCG depended upon single nucleotide polymorphisms (SNP) in genes implicated as important for cow fertility. We also evaluated whether a SNP in a gene important for cellular resistance to heat stress (HSPA1L, a member of the heat shock protein 70 family) is important for embryonic resistance to elevated temperature. Major conclusions, solutions & achievements. Results confirmed that elevated temperature increases ROS production by the oocyte and embryo and that melatonin decreases ROS. Melatonin reduced, but did not completely block, damaging effects of heat shock on the oocyte and had no effect on development of the embryo. Melatonin was protective to the oocyte at 0.1-1 μM, a concentration too high to be achieved in cows. It was concluded that melatonin is unlikely to be a useful molecule for increasing fertility of heat-stressed cows. Treatment with hCG at day 5 after breeding increased first-service pregnancy rate for primiparous cows but not for multiparous cows. Thus, hCG could be useful for increasing fertility in first-parity cows. The effectiveness of hCG depended upon genotype for a SNP in COQ9, a gene encoding for a mitochondrial-function protein. This result points the way to future efforts to use genetic information to identify populations of cows for which hormone treatments will be effective or ineffective. The SNP in HSPA1L was related to embryonic survival after heat shock. Perhaps, genetic selection for mutations that increase cellular resistance to heat shock could be employed to reduce effects of heat stress on fertility. Implications, both scientific and agricultural. This project has resulted in abandonment of one possible approach to improve fertility of the heat-stressed cow (melatonin therapy) while also leading to a method for improving fertility of primiparous cows exposed to heat stress (hCG treatment) that can be implemented on farms today. Genetic studies have pointed the way to using genetic information to 1) tailor hormonal treatments to cow populations likely to respond favorably and 2) select animals whose embryos have superior resistance to elevated body temperatures.

Infectious disease is one of the most serious causes of economic loss in all sectors of aquaculture. There is a critical need to understand the molecular basis for protection against infectious disease so that safer, more reliable and more cost-effective strategies can be designed for their control. As part of this effort, the major goal of our BARD project was to determine the importance of endobiotics as a defense against protozoan ectoparasites in fish. Endobiotics, or antimicrobial polypeptides, are peptides and small proteins that are increasingly recognized as having a vital role in the innate defense of virtually all animals. One objective of our BARD project was to determine the antiparasitic potency of one specific group of endobiotics that were isolated from hybrid striped bass (Morone saxatilis x M chrysops). We found that these endobiotics, which we had previously named histone-like proteins (HLPs), exhibited potent activity against Amyloodinium and that the putative levels of HLPs in the skin were well within the levels that we found to be lethal to the parasite in vitro. We also found evidence for the presence of similar antibiotics in sea bream (Sparus aurata) and Mediterranean sea bass (Dicentrarchus labrax). We also examined the effect of chronic stress on the expression of HLP in fish and found that HLP levels were dramatically decreased after only one week of a crowding/high ammonia sublethal stress. We also began to explore the feasibility of upregulating endobiotics via immunostimulation. However, we did not pursue this objective as fully as we originally intended because we spent a much larger effort than originally anticipated on the last objective, the attempted isolation of novel endobiotics from hybrid striped bass. In this regard, we purified and identified four new peptide endobiotics. These endobiotics, which we have named piscidins (from "Pisces" meaning fish), have potent, broad-spectrum activity against a number of both fish and human pathogens. This includes not only parasites but also bacteria. We also demonstrated that these peptides are present in the mast cell. This was the first time that the mast cell, the most common tissue granulocyte in vertebrates, was shown to possess any type of endobiotic. This finding has important implications in explaining the possible function of mast cells in the immune response of vertebrates. In summary, the research we have accomplished in this BARD project has demonstrated that endobiotics in fish have potent activity against many serious pathogens in aquaculture and that there is considerable potential to use these compounds as stress indicators in aquaculture. There is also considerable potential to use some of these compounds in other areas of medicine, including treatment of serious infectious diseases of humans and animals.