Identifying selection strategies based on the practices and preferences of small ruminant farmers to improve the sustainability of their breeding systems.

Small ruminant farming is of socio-economic and environmental importance to many rural communities around the world. The SMARTER H2020 project aims to redefine genetic selection criteria to increase the sustainability of the sector. The objective of this study was to analyse the selection and breeding management practices of small ruminant producers and breeders, linked with socio-technical elements that shape them. The study is based on farm surveys using semi-structured interviews conducted in five countries (France, Spain, Italy, Greece, and Uruguay) across 272 producers and breeders of 13 sheep and goat breeds, and 15 breed × system combinations. The information was collected in four sections. The first and second sections dealt with general elements of structure and management of the system and the flock/herd. The third section focused on selection and breeding management practices: criteria for culling and replacement of females, selection criteria for males, use of estimated breeding values and global indexes, and preferences for indexing new traits to increase the sustainability of their system. The fourth section aimed to collect socio-technical information. We used a data abstraction method to standardise the representation of these data. A mixed data factor analysis followed by a hierarchical ascending classification allowed the characterisation of three profiles of selection and breeding management: (1) a profile of producers (n = 93) of small flocks/herds, with little knowledge or use of genetic selection and improvement tools (selection index, artificial insemination, performance recording); these farmers do not feel that new traits are needed to improve the sustainability of their system. (2) a profile of producers (n = 34) of multibreed flocks/herds that rely significantly on grazing; they are familiar with genetic tools, they currently use AI; they would like the indexes to include more health and robustness characteristics, to make their animals more resistant and to increase the sustainability of their system. And (3) a profile of producers or breeders (n = 145) of large flocks/herds, with specific culling criteria; these farmers are satisfied with the current indexes to maintain the sustainability of their system. These results are elements that can be used by private breeding companies and associations to support the evolution of selection objectives to increase the resilience of animals and to improve the sustainability of the small ruminant breeding systems.

Inflammation and thrombo-occlusive vessel signalling in benign atrophic papulosis (Köhlmeier-Degos disease).

BACKGROUND: Although the merely cutaneous, benign form of the extremely rare disease atrophic papulosis (Köhlmeier-Degos disease) may occasionally develop into the systemic, malignant form with time, it is unclear whether it exhibits any systemic characteristics. OBJECTIVE: To determine whether benign atrophic papulosis exhibits inflammatory and thrombo-occlusive signals and to classify it according to the Chapel-Hill classification of vasculitis. METHODS: In a monocentric, controlled study, levels of cytokines (IL-1ß, IL-6, IL-8, IFNγ, MCP-1, VEGF, TNFα, TGF-ß1), antiphospholipid antibodies (cardiolipin IgG/A/M, cardiolipin IgG, cardiolipin IgM, ß2-glycoprotein IgG/A/M, phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol, phosphatidyl ethanolamine and sphingomyelin A), antibodies against proteinase-3 IgG and myeloperoxidase IgG, antinuclear antibodies and extractable nuclear antigen were assessed in blood samples of six benign atrophic papulosis patients and six age- and sex-matched healthy controls. RESULTS: IL-8 was only detectable in patients' serum. VEGF was reduced and cardiolipin IgG/A/M and ß2-glycoprotein antibodies were increased in the patients' group. ANA were only detected in three patients, and ENA were negative throughout. No differences were detected between the other investigated markers. CONCLUSIONS: This is the first study evaluating systemic inflammatory and thrombo-occlusive vessel signalling in benign atrophic papulosis and provides evidence of a non-antineutrophil cytoplasmatic antibodies immune-complex small vessel vasculitis according to the Chapel-Hill classification. These findings corroborate its systemic character despite the apparent missing involvement of systemic organs.

Atrophic papulosis (Köhlmeier-Degos disease) revisited: a cross-sectional study on 105 patients.

BACKGROUND: Atrophic papulosis is a very rare vascular disease of unknown pathogenesis, mostly described by case reports. OBJECTIVE: To assess demographic data and prognosis in patients with atrophic papulosis. METHODS: A single-centre study was performed on a series of 105 patients with atrophic papulosis, diagnosed 2000-2021. Patients were referred and diagnosed at the evaluation centre and patients' clinical data were provided by the Degos Support Network and evaluated by the authors for confirming the diagnosis of skin lesions and fulfilling the diagnostic criteria for a malignant subset. A unique set of variables were collected from all patients. RESULTS: The mean age of disease onset was 33.3 ± 18.3 years and the male-to-female ratio was 1:1.6. The family history rate was 8.1%. The classification into a benign, merely cutaneous disease (benign atrophic papulosis), and malignant atrophic papulosis, associating cutaneous and visceral lesions was confirmed due to their striking prognostic difference. Benign atrophic papulosis was detected in 41% of the patients with no deaths occurring throughout the follow-up period (median 3.00 years; range 0.13-23). Malignant atrophic papulosis was reported in 59% of patients with 47.5% multisystemic involvement and a median skin lesion onset to systemic symptoms duration of 0.54 years (-6 to 20). The gastrointestinal tract and central nervous system were equally involved; however, the neurological involvement-caused death rate was slightly higher. The disease-specific mortality rate of malignant atrophic papulosis was 22.6%. CONCLUSIONS: Atrophic papulosis presents with a striking prognostic difference of benign - merely cutaneous - involvement or quickly developing - into less than 1 year - malignant subset, associating cutaneous and visceral lesions and multiorgan involvement in 1/2 of the patients, which leads to premature, disease-specific mortality in 1/4 of the cases. Central nervous system and gastrointestinal tract complications are the major reasons for disease-specific death. Over the years, the diagnosis of severe nervous system involvement has become more common.

Chios ram testicular blood flow and echotexture changes depending on age, season and ejaculation process.

The present study aimed to investigate the impact of age, season and ejaculation on ram testicular blood flow and echotexture. The survey was conducted biweekly on 7 Chios rams for one year, including breeding and non-breeding periods. The rams were divided into 2 age groups: 3 rams 2-6 years old (mature) and 4 rams 9-13 years old (old). Hemodynamic indices [Pulsatility index (PI), Resistive index (RI), End-diastolic velocity (EDV), testicular artery Diameter (D), Time-averaged maximum velocity (TAVM), Blood flow volume (BFV)] and echotexture parameters [Mean value (MV), Contrast (Con), Gray value distribution (GVD), Run length distribution (RunLD), Long run emphasis (LRunEm), Entropy (Ent), Correlation (Cor), Standard deviation (StD), Gray variance (GV) and Gradient mean value (GMV)] were evaluated in each testis before and after ejaculation. Ejaculation did not affect testes blood flow or echotexture (p>0.05). PI and RI were higher in the breeding period compared to the non-breeding period, for both testes (p⟨0.001). Left testis GV and Cor before ejaculation were lower (p=0.01) and higher (p=0.03), respectively, in the breeding compared to the non-breeding period. Left testis D (p=0.005) and BFV (p⟨0.001) were higher in old compared to mature rams after ejaculation. Right testis Con (p=0.03) and Cor (p=0.05) before ejaculation were higher in old rams, whereas right testis Ent after ejaculation was higher in mature rams (p=0.05). In conclusion, testicular blood flow and echotexture are affected by season and ram age, but not by ejaculation.

Effect of astaxanthin in extenders on sperm quality and functional variables of frozen-thawed boar semen.

The aim of the study was to determine whether the presence of astaxanthin (ASX) protects boar spermatozoa against damage related to cryopreservation. Pooled ejaculates extended in Beltsville Thawing Solution (BTS) were used. Three experiments were conducted: 1) sperm samples were pre-incubated overnight (17 °C) with ASX (0, 0.5, 5, 15 µM) prior to freezing and then frozen using cooling and thawing extenders supplemented with ASX (0, 0.5, 5, 15 µM); 2) sperm samples were treated with ASX (0, 0.5, 5, 15 µM) only during overnight pre-incubation (17 °C) prior to cryopreservation; and 3) a thawing extender was supplemented with ASX (0, 0.5, 5, 15 µM). The groups were as follows: control (C; no treatment), ASX 1 (0.5 µM), ASX 2 (5 µM) and ASX 3 (15 µM). Total (TM) and progressive (PM) motility was analyzed using CASA, while sperm viability, reactive oxygen species generation, lipid peroxidation and apoptoticlike changes were analyzed using flow cytometry. Sperm variables were evaluated prior to freezing as well as 30 and 150 min after thawing. In Experiment 1, the values of TM and sperm viability post-thaw were less in the ASX 3 than C group. In Experiment 2, there was no effect of ASX on any of the sperm variables evaluated, while in Experiment 3, apoptotic-like changes were less in the ASX 1 than C group. In conclusion, there was a subtle beneficial effect on cryopreserved boar spermatozoa after addition of ASX to thawing media.

Invited review: Current production trends, farm structures, and economics of the dairy sheep and goat sectors.

Dairy small ruminants account for approximately 21% of all sheep and goats in the world, produce around 3.5% of the world's milk, and are mainly located in subtropical-temperate areas of Asia, Europe, and Africa. Dairy sheep are concentrated around the Mediterranean and Black Sea regions, where their dairy products are typical ingredients of the human diet. Dairy goats are concentrated in low-income, food-deficit countries of the Indian subcontinent, where their products are a key food source, but are also present in high-income, technologically developed countries. This review evaluates the status of the dairy sheep and goat sectors in the world, with special focus on the commercially and technically developed industries in France, Greece, Italy, and Spain (FGIS). Dairy small ruminants account for a minor part of the total agricultural output in France, Italy, and Spain (0.9 to 1.8%) and a larger part in Greece (8.8%). In FGIS, the dairy sheep industry is based on local breeds and crossbreeds raised under semi-intensive and intensive systems and is concentrated in a few regions in these countries. Average flock size varies from small to medium (140 to 333 ewes/farm), and milk yield from low to medium (85 to 216 L/ewe), showing substantial room for improvement. Most sheep milk is sold to industries and processed into traditional cheese types, many of which are Protected Denomination of Origin (PDO) cheeses for gourmet and export markets (e.g., Pecorino, Manchego, and Roquefort). By comparing break-even milk price among FGIS countries, we observed the following: (1) most Greek and French dairy sheep farms were unprofitable, with the exception of the intensive Chios farms of Greece; (2) milk price was aligned with cost of production in Italy; and (3) profitable farms coexisted with unprofitable farms in Spain. In FGIS, dairy goat production is based on local breeds raised under more extensive systems than sheep. Compared with sheep, average dairy goat herds are smaller (36 to 190 does/farm) but milk yield is greater (153 to 589 L/doe), showing room for improvement. Goat milk is mainly processed on-farm into dairy products for national markets, but some PDO goat milk cheeses (e.g., Murcia al Vino) are exported. Processed goat milk is sold for local human consumption or dehydrated for export. Mixed sheep-goat (e.g., Feta) and cow-sheep-goat milk cheeses are common in many countries. Strategies to improve the dairy sheep and goat sectors in these 4 countries are proposed and discussed.

Defining the breeding goal for a sheep breed including production and functional traits using market data.

In this study, the economic values for production and functional traits of dairy sheep are estimated through the application of a profit function model using farm-level technical and economic data. The traits incorporated in the model were milk production, prolificacy, fertility, milking speed, longevity and mastitis occurrence. The economic values for these traits were derived as the approximate partial derivative of the specified profit function. A sensitivity analysis was also conducted in order to examine how potential changes in input and output prices would affect the breeding goal. The estimated economic values of the traits revealed their economic impact on the definition of the breeding goal for the specified production system. Milk production and fertility had the highest economic values (€40.30 and €20.28 per standard genetic deviation (SDa)), while, mastitis only had a low negative value of -0.57 €/SDa. Therefore, breeding for clinical mastitis will have a minor impact on farm profitability because it affects a small proportion of the flock and has low additive variance. The production traits, which include milk production, prolificacy and milking speed, contributed most to the breeding goal (70.0%), but functional traits still had a considerable share (30.0%). The results of this study highlight the importance of the knowledge of economic values of traits in the design of a breeding program. It is also suggested that the production and functional traits under consideration can be categorized as those which can be efficiently treated through genetic improvement (e.g. milk production and fertility) while others would be better dealt with through managerial interventions (e.g. mastitis occurrence). Also, sub-clinical mastitis that affects a higher proportion of flocks could have a higher contribution to breeding goals.

Effect of colostrum quantity and quality on neonatal calf diarrhoea due to Cryptosporidium spp. infection.

This study was conducted to assess the effect of colostrum quality and quantity on Cryptosporidium spp. calf diarrhoea in an intensive dairy cattle farm in Greece. Faecal samples were collected from 100 dairy calves randomly selected and born during all 4 seasons (March 2015 to May 2016) of the year. In total, 71% of the selected calves were positive for Cryptosporidium spp. oocysts. The statistical analysis revealed influence of colostrum quality on faecal score. Linear regression showed that the colostrum quantity during the first day of life was negatively associated with the number of Cryptosporidium spp. oocysts in faeces. During multivariable analysis, the variables representing the quality of colostrum and the season of the calf's birth were identified as confounders. Cryptosporidium spp. is a common pathogen participating in neonatal calf diarrhoea. Colostrum management and season influence the number of Cryprosporidium spp. oocysts and faecal consistency. The above findings demonstrate novel risk factors that should be included in the strategic approaches to control cryptosporidiosis in newborn calves.

Benign atrophic papulosis (Köhlmeier-Degos disease): the wedge-shaped dermal necrosis can resolve with time.

BACKGROUND: Atrophic papulosis is a rare thrombo-occlusive disease, characterized by the appearance of multiple atrophic porcelain-white skin papules, with a surrounding erythematous rim, which are histologically consisting of wedge-shaped necrosis of the dermis. OBJECTIVE: It consists of two variants: (i) the benign atrophic papulosis (BAP) only involving the skin and (ii) the malignant atrophic papulosis (MAP) also involving several internal organs with a cumulative five-year survival rate of approx. 55%. While the probability of only having a BAP at onset is approximately 70%, increasing to 97% after 7 years of monosymptomatic cutaneous course, no close long-term follow-up of the development of the skin lesions has been reported. METHODS: We present a precise visual documentation of the evolution of the disseminated skin lesions in a female patient with BAP spanning over two decades. RESULTS: A considerable improvement and/or clinical resolution of the majority of the lesions disputing the scarring character of the atrophic porcelain-white skin papules has been detected. CONCLUSION: BAP not only exhibits an excellent prognosis, but resolution of lesions can also occur after a considerable period of time.

Protective effect of crocetin on bovine spermatozoa against oxidative stress during in vitro fertilization.

Oxidative stress is one of the major factors that contribute to poor semen quality and low rates of in vitro fertilization. Crocetin, a main constituent of saffron (Crocus sativus L.) possesses potent antioxidant activity, by scavenging reactive oxygen species (ROS) and/or enhancing the activity of intracellular antioxidant enzymes. The aim of this study was to investigate, for the first time, the effect of crocetin on the quality characteristics of bull spermatozoa and fertilization rate. For this reason, frozen/thawed bovine spermatozoa were incubated with crocetin (1, 2.5, and 5 µm), for 120 or 240 min, in the presence of a negative control, and evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. In order to evaluate the impact of crocetin on cleavage and blastocyst rate, the compound was added in the IVF medium at the previously identified optimal concentration (2.5 µm). The results indicate that incubation of spermatozoa with 2.5 µm of crocetin resulted in a statistically significant lower production of superoxide anion and hydrogen peroxide, lower lipid peroxidation, and in better maintenance of motility parameters, viability, and acrosomal integrity, with a very small number of cells with DNA fragmentation, compared to the other groups (p < 0.05). The presence of crocetin (2.5 µm) in the fertilization medium also resulted in a significant increase in acrosome-reacted spermatozoa and blastocyst production, compared to the control group (p < 0.01). These data indicate that crocetin (2.5 µm) positively affects bovine sperm quality characteristics during a 240-min incubation and improves its fertilizing ability, directly and/or indirectly, by regulating ROS concentration and lipid peroxidation.

Antioxidant effect of crocin on bovine sperm quality and in vitro fertilization.

Reactive oxygen species (ROS) production above critical levels affects the genetic and functional integrity of spermatozoa by causing oxidative stress. Spermatozoa are susceptible to oxidative stress in terms of motility and fertilization capacity. Crocin (crocetin di-gentiobiose ester), a main constituent of Crocus Sativus L. (saffron), is known for its antioxidant activity by scavenging ROS, especially superoxide anion. The aim of the present study is to evaluate the effect of crocin on the quality characteristics of spermatozoa and fertilization rate. Frozen-thawed and washed spermatozoa from four different bulls were incubated with three different concentrations of crocin (0.5, 1, and 2 mM), for 120 and 240 minutes, in the presence of a negative control, and were evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. The most potent concentration of crocin (1 mM) was also added in the fertilization medium to test its impact on fertilization outcome. The results indicate that the incubation of spermatozoa with 1 mM of crocin resulted in a statistically significant lower production of ROS, lower lipid peroxidation and in better maintenance of motility, viability, and acrosomal integrity, with a very small number of fragmented cells, compared to the control and the other treated groups (P < 0.05). Crocin concentration of 1 mM resulted in a significant increase of blastocyst rate, compared to the control group (P < 0.01). These data indicate that crocin (1 mM) improves bovine sperm quality and its fertilization capability, directly and/or indirectly, by modulating ROS concentration.

Reproductive performance of sows was improved by administration of a sporing bacillary probiotic (Bacillus subtilis C-3102).

This field study assessed the efficacy of a probiotic based on viable spores of Bacillus subtilis C-3102 (Calsporin; Calpis Co. Ltd., Japan) on the health status and productivity of sows and their litters through 2 full, sequential reproductive cycles from service of the first cycle to weaning of the second cycle. Fifty-six sows were allocated to 2 experimental groups, an untreated control (T1) group and a probiotic-treated (T2) group that received the same basal feed as the T1 group plus the probiotic at an approximate allowance of 30 g/t of feed (3 × 10(5) cfu/g). The offspring of T1 and T2 sows were offered basal and T2 creep feed (3 × 10(5) cfu/g), respectively. Health and zootechnical parameters of sows and piglets were recorded. Feeding the probiotic to sows and piglets resulted in significant benefits, observed in both cycles: 1) improved sow body condition during pregnancy (P < 0.05), 2) increased sow feed consumption, 3) reduced sow weight loss during lactation (P < 0.05), 4) reduced sow weaning-estrus interval (P < 0.05), and 5) higher BW of piglets at weaning (P < 0.05). Additionally, a significant (P < 0.05) improvement in piglet birth weight and in the number of piglets weaned was observed in the second cycle of T2 sows, while a significant improvement of mean daily gain of piglets from birth to weaning was observed in the first cycle of T2 sows. Microbiological examination of fecal samples showed that probiotic treatment significantly reduced both Escherichia coli and Clostridium spp. in piglet feces, particularly during the second cycle. The data suggested that continuous feed supplementation with the probiotic is beneficial for both sows and piglets, since zootechnical benefits were observed in both cycles.

Effects of sexual maturation and Salmonella infection on the expression of avian ß-defensin genes in the chicken testis.

Rooster infertility is a major concern in the poultry industry and protection of the male reproductive organs from pathogens is an essential aspect of reproductive physiology. During the last years, research on antimicrobial protection has elucidated the critical role of the antimicrobial peptides avian ß-defensins (AvBDs) in the innate immunity in chickens. AvBDs have been reported to be expressed in the hen reproductive organs, providing protection against microbial pathogens including Salmonella Enteritidis (SE). However, mechanisms of antimicrobial protection of rooster reproductive organs and especially the testis, mediated by AvBDs are poorly understood. The aim of this study was to investigate the expression of the complete family of the 14 AvBD genes, in the rooster testis in vivo, to determine whether sexual maturation affects their testicular mRNA abundance and to investigate whether SE infection alters their expression. Expression analysis revealed that 9 members of the AvBD family, namely AvBD1, 2, 4, 5, 6, 9, 10, 12 and 14 were expressed in the testis. Quantitative real-time PCR analysis revealed that the mRNA abundance of three AvBDs was up regulated and of three AvBDs was down regulated with respect to sexual maturation. In addition, SE infection resulted in a significant induction of AvBD4, 10, 12 and 14 in the testis of sexually mature roosters. These findings provide strong evidence to suggest that an AvBD-mediated immune response mechanism exists in the rooster testis providing protection against bacterial pathogens including Salmonella species.

Malignant and benign forms of atrophic papulosis (Köhlmeier-Degos disease): systemic involvement determines the prognosis.

BACKGROUND: Atrophic papulosis (Köhlmeier-Degos disease) is a rare disease of unknown aetiology. The cutaneous signs--papular skin lesions with central porcelain-white atrophy and surrounding telangiectatic rim--are almost pathognomonic. Extracutaneous, systemic involvement includes multiple limited infarcts of the gastrointestinal system, central nervous system and other organs. OBJECTIVES: To assess prospectively the demographics, epidemiological data and prognosis of patients with atrophic papulosis evaluated in a single centre. METHODS: A prospective, single-centre, cohort study at diagnosis was performed on a series of 39 patients with atrophic papulosis, first seen between 2000 and 2007 and evaluated up to 2012. RESULTS: The occurrence of cutaneous lesions defined the onset of disease in all cases. The mean age of onset was 35.4 ± 12.3 years and the male-to-female ratio was 1 : 1.4. In total, 9% of patients reported familial occurrence. Extracutaneous (systemic) signs were recorded in 29% of the patients, whereas the median time for development of systemic manifestations was 1 year (0.03-0.97 quantiles: 0-7 years) after the occurrence of cutaneous lesions. The prognosis was determined mainly by the presence of systemic involvement. 73% of the patients with systemic manifestations (73% developed intestinal perforation) died, while none of the patients with only cutaneous disease had a lethal outcome. The cumulative 5-year survival rate in patients with systemic disease was 54.5%. CONCLUSIONS: Atrophic papulosis, previously called malignant atrophic papulosis, should be classified into a malignant, systemic form and a benign, cutaneous one, the latter being more common. The probability of having a benign form of the disease at onset is approximately 70%, increasing to 97% after 7 years of monosymptomatic cutaneous course.

Technical indicators of economic performance in dairy sheep farming.

In this study, the level of technical efficiency of 58 sheep farms rearing the Chios breed in Greece was measured through the application of the stochastic frontier analysis method. A Translog stochastic frontier production function was estimated using farm accounting data of Chios sheep farms and the impact of various socio-demographic and biophysical factors on the estimated efficiency of the farms was evaluated. The farms were classified into efficiency groups on the basis of the estimated level of efficiency and a technical and economic descriptive analysis was applied in order to illustrate an indicative picture of their structure and productivity. The results of the stochastic frontier model indicate that there are substantial production inefficiencies among the Chios sheep farms and that these farms could increase their production through the improvement of technical efficiency, whereas the results of the inefficiency effects model reveal that the farm-specific explanatory factors can partly explain the observed efficiency differentials. The measurement of technical inefficiency and the detection of its determinants can be used to form the basis of policy recommendations that could contribute to the development of the sector.

Changes in the expression of Toll-like receptors in the chicken testis during sexual maturation and Salmonella infection.

Rooster infertility is a major concern in the poultry industry and chicken male reproductive organs are the infectious tissues of various pathogenic microorganisms. Protection of the chicken male reproductive organs from pathogens is therefore an essential aspect of reproductive physiology. Recently Toll-like receptors (TLRs) have been identified as one of the key components of innate immunity in vertebrate species and have been reported to be expressed in the reproductive organs in various female species, including the chicken. However, mechanisms of antimicrobial protection of male reproductive organs mediated by TLRs are poorly understood. The objectives of this study were to determine the expression profile of the entire family of the ten chicken TLR genes in the chicken testis, to investigate whether sexual maturation affects their testicular mRNA abundance and to determine the changes in their expression levels in response to Salmonella enteritidis (SE) infection. RNA was extracted from the testis of healthy pre-pubertal, sexually mature and aged birds, and from sexually mature SE infected birds. RT-PCR analysis revealed that all TLRs, apart from TLR1-1 (TLR6), were expressed in the chicken testis. Quantitative real-time PCR analysis revealed that the testicular mRNA abundance of certain TLRs was developmentally regulated with respect to sexual maturation, while SE infection resulted in a significant induction of TLR2-1, 4, 5, 15 and 21 in the testis of sexually mature birds compared, to healthy birds of the same age. These findings provide strong evidence to suggest a key role of TLRs in the innate immune responses of chicken testis against Salmonella colonization.

Effects of sexual maturation and Salmonella infection on the expression of Toll-like receptors in the chicken vagina.

Toll-like receptors (TLRs) are a critical component of the innate immune response in many vertebrates, including avian species. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles and in the chicken ovary in vivo, as well as the changes in their expression in response to lipopolysaccharide or Salmonella enteritidis (SE) infection, have broad implications for reproductive physiology and for the prevention of transmission of zoonotic diseases to humans through the consumption of contaminated poultry eggs. Because the main route of egg contamination is from infection of the oviduct and mainly from the vagina, the aim of this study was to investigate the expression of the ten cTLRs identified to date in the chicken oviduct in vivo, to determine whether sexual maturation affects their mRNA abundance and to investigate whether SE infection alters the expression of TLRs in the chicken vagina. RNA was extracted from the vagina of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all types of cTLRs apart from TLR1-1 were expressed in the vagina of sexually mature birds. Quantitative real-time PCR analysis revealed that the mRNA abundance of TLR2-1, 2-2 and 4 differ with respect to sexual maturation in the chicken vagina. SE infection resulted in a significant induction of TLR5 and 15 in the vagina of sexually mature birds, and in a significant induction of TLR2-1, 4 and 15 in the vagina of aged birds, while a significant down-regulation was observed for TLR7 in the vagina of sexually mature birds. These findings suggest that a TLR mediated immune response mechanism exists in the chicken vagina, playing a crucial role in preventing microbial pathogens from being incorporated into newly forming eggs.

Transcriptional profiling of Toll-like receptors in chicken embryos and in the ovary during sexual maturation and in response to Salmonella enteritidis infection.

One of the key members of the innate immune system in many vertebrate species is the family of Toll-like receptors (TLRs). These molecules, which initiate the innate immune response and mount an anti-microbial response in both vertebrates and invertebrates, have recently been identified in the chicken genome. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles during follicular growth and in response to lipopolysaccharide (LPS) infection are very important for reproductive physiology due to the transovarian transmission of Salmonella enteritidis (SE) in laying hens. The aim of this study was to investigate the expression of the ten cTLRs identified to date, in the chicken ovary in vivo and embryos during early embryonic development, to investigate whether sexual maturation affects their ovarian mRNA abundance and to investigate the transcriptional changes of TLRs in the chicken ovary in response to SE infection. RNA was extracted from embryos from day 3 to day 10 of embryonic development as well as from the ovaries of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all TLRs apart from TLRs 1-1 and 2-2 were expressed in the ovary of sexually mature chickens, while all TLRs apart from TLR1-1 were expressed in the chicken embryos during embryonic development. Quantitative real-time PCR analysis revealed that the ovarian mRNA abundance of TLRs differ with respect to sexual maturation. SE infection resulted in a significant induction of TLR4, and 15 in the ovary of sexual mature birds, and in a significant induction of TLR15 in the ovary of aged birds, while a significant down-regulation was observed for TLR3 in the ovary of aged birds. These findings suggest that a TLR-mediated immune response mechanism exists in the chicken ovary.

Reduction of cell mediated immunity in patients with genital warts of long duration.

Cell mediated immunity was studied by a leucocyte migration inhibition assay and by tuberculin and dinitrochlorobenzene skin tests in 30 patients with recurrent genital warts and in 34 healthy people (with no history of genital warts) who served as controls. Migration inhibition was significantly less in patients suffering from recurrences for more than one year than in controls (p less than 0.001). Dinitrochlorobenzene and tuberculin sensitivity were also found to be impaired in those with infection of long duration (p less than 0.001).

Studies on bovine herpesviruses. Part 1. Isolation and characterization of viruses isolated from the genital tract of cattle.

Herpesviruses, previously isolated from cattle (Theodoridis, 1978), were further studied and provisionally placed in the bovid herpesvirus 4 (BHV-4) group. Major differences were found between IBR-IPV (BHV-1) and BHV-4 virus strains. In MDBK cells, all BHV-4 strains started growing at the edges of the culture, the process progressing slowly until destruction of the cells was complete by the 10th day. BHV-4 strains failed to induce neutralizing antibodies in cattle, goats and rabbits. Only the addition of mineral oil adjuvant induced neutralizing and complement fixing antibodies in goats. BHV-1 strains, in contrast, produced very potent antisera in all these systems. Cross-neutralization tests indicated the existence of 2 distinct serological groups representing BHV-1 and BHV-4. The BHV-4 strains appear to be interrelated and they could not be grouped. A BHV-1 strain showed fixation of complement with the antisera of 6 BHV-4 strains. Electron micrographs showed an accumulation of nucleocapsids in the cytoplasm and an early release of virus particles due to cell destruction. Variation in incubation temperature had a significant effect on the particle formation. At lower temperatures, the number of enveloped particles in the cytoplasm increased. On the basis of the characteristics uncovered in this study, it is possible that all the BHV-4 strains represent one and the same virus which has undergone certain biological changes, thus illustrating a phenomenon which appears to be a characteristic of the herpesviruses.