Enhancement of the interferon gamma assay to detect paratuberculosis using interleukin-7 and interleukin-12 potentiation

A limitation to the widespread application of interferon gamma (IFN-γ) response assays has been logistical difficulties, as they must be performed within hours of blood collection. Detection of specific IFN-γ responses to Mycobacterium avium subspecies paratuberculosis (MAP) as part of the cell-mediated immune response of ruminants with Johne's disease could aid in diagnosis and control programs. In this study, a modified protocol was developed in which cultures were supplemented with interleukin (IL)-7, a survival factor required to maintain resting T cells, alone or in combination with IL-12 to potentiate IFN-γ responses and extend blood storage time. The combination of IL-7 and IL-12 was synergistic, giving IFN-γ responses greater than with IL-12 alone, for sheep blood stored up to 2 days. In a cohort of naturally infected sheep, the same number of animals was identified as test positive using the modified assay (performed after 2 days with IL-7/IL-12 supplementation) as the standard IFN-γ assay performed on the day of blood collection. The modified assay offered greatest advantage in the detection of early stages of paratuberculosis infection, for sheep with low grade and paucibacillary lesions, and at early time points post-infection. The potentiation protocol allowed for practical shipment of blood samples from farm to laboratory, extending permissible transit times and applicability of IFN-γ testing to detect Johne's disease.     

Effects of α-tocopherol and Ascorbic Acid on Equine Semen Quality after Cryopreservation

This study investigated the effects of ascorbic acid and α-tocopherol supplementation on semen quality parameters of equine thawed-frozen semen. Semen was divided in seven different treatments in a final concentration of 100 × 10⁶ sperm/mL by using Gent extender containing no supplements (control) and the following supplements with three different concentrations: α-tocopherol (0.5, 1, and 2 mM) and ascorbic acid (0.45, 0.9, and 1.8 g/L). After thawing, all samples were maintained at 37°C, while analyses were performed at 0, 60, and 120 minutes. Evaluation of viability and acrosome status (using Pisum sativum agglutinin conjugated to fluorescein isothiocyanate and propidium iodide), mitochondrial membrane potential (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine [JC-1]), membrane lipid peroxidation (LPO; C₁₁-BODIPY^581/591), and stability of the plasmatic membrane (merocyanine 540 and Yo-Pro-1) of each sample was determined by flow cytometry. Relative to the control group, supplementation with α-tocopherol improved (P ≤ .05) postthaw membrane LPO, yet the higher concentrations of ascorbic acid (0.9 and 1.8 g/L, respectively) showed a negative effect on membrane LPO. Neither antioxidant significantly increased (P > .05) the acrosome integrity and mitochondrial membrane potential of frozen-thawed spermatozoa, although supplementation with α-tocopherol and ascorbic acid (0.9 and 1.8 g/L, respectively) had a positive effect on membrane integrity and stability (P ≤ .05). For all semen parameters, the lower concentration of ascorbic acid (0.45 g/L) did not show significant differences (P > .05) compared with the control. In conclusion, α-tocopherol seems to be an efficient antioxidant for reducing the oxidative stress provoked by cryopreservation, decreasing lipid peroxidation on equine spermatozoa.     

Seroprevalence of Antibodies to Chlamydophila psittaci in Zoo Workers in Brazil

To evaluate the prevalence of antibodies to Chlamydophila psittaci 364 serum samples were collected from veterinarians, biologists, animal scientists, veterinary students, animal keepers and others employees in 20 zoos, and from veterinary practitioners in 10 Brazilian states. Subjects ranged from 15 to 64 years of age, with 268 (74%) males and 96 (26%) females. Chlamydial antibodies were determined by the complement fixation test (CFT) and specific anti‐C. psittaci IgG antibodies were determined by the microimmunoflurescence (MIF) test. Complement fixation test showed 23.9% (87/364) and MIF test showed 4.7% (17/364) positive serum samples. Titres ranged from 16 to 256 in both assays, demonstrating evidence of recent or current infection. Although chlamydial antibodies were detected in workers of seventeen zoos, MIF test only detected specific C. psittaci antibodies in seven of them. Previous psittacosis infection was suspected in eight workers of two zoos, five of whom reported having pneumonia, while employed at the zoos. However, diagnosis was not established in any of these cases in the past. Results indicated the occurrence of infection and previous contact of Brazilian zoo workers with C. psittaci, as well as the zoonotic potential of psittacosis in this risk population. Other studies are necessary to evaluate the risk factors of infection in this population. This seroepidemiological survey confirmed the need to adopt preventive measures to control avian chlamydiosis and protect the health of zoo workers in the country.     

Microvascular development and growth of uterine tissue during the estrous cycle in mares

OBJECTIVE: To document uterine growth and microvascular development in the endometrium of uteri with differing degrees of fibrosis as well as uterine growth throughout the estrous cycle of mares. ANIMALS: 30 mares. PROCEDURE: Uterine tissue was obtained during the breeding season from a slaughter facility. Stage of estrous cycle of the mares was assessed on the basis of ovarian structures and plasma progesterone concentrations. Endometrium was characterized by use of light microscopy, and blood vessel walls were marked by histochemical techniques. Microvascular development was evaluated by a computerized image analysis system. Growth of uterine tissue was based on cellular content of DNA and RNA, RNA:DNA, and protein:DNA. RESULTS: Significant differences in vascular density were not observed in the endometrium of uteri obtained from mares euthanatized during the follicular or luteal phase of the estrous cycle, regardless of whether endometrial classification of degree of fibrosis was considered. There was a 3-fold increase in amount of DNA and RNA of endometrial cells in the follicular phase when compared to myometrium. Hypertrophy of endometrial tissue during the luteal phase was reflected by a significant increase in cell protein content and protein:DNA. CONCLUSIONS AND CLINICAL RELEVANCE: Endometrial growth of vascular tissues during the estrous cycle may be coordinated with development of nonvascular tissue. Estrogen and progesterone may play a role in regulation of uterine growth and angiogenesis.     

Quantitative and descriptive histological aspects of jaguar (Panthera onca Linnaeus, 1758) ear skin as a step towards formation of biobanks

Skin of mammals vulnerable to extinction, such as the jaguar, is used as a source of material in conservation strategies. The composition of skin is not uniform among species, and the ability to distinguish similarities in skin morphology in animal groups is fundamental in the application of skin tissue for use in biobanks. The aim of our study was to evaluate the structure, composition and capacity for culture of ear skin from the yellow and black jaguars. Both qualitative and quantitative methods were used, focusing on skin thickness, cell quantification and distribution, collagen density, proliferative activity and viability. Histomorphometrical study of the skin showed a total thickness of 273.2 and 274.6 µm for the yellow and black jaguars, respectively. Melanocytes and fibroblasts were, respectively, 9.7 and 23.0 for the yellow jaguar and 11.3 and 26.8 for the black jaguar. A collagen density of 67.0% and 49.0% was observed for yellow and black jaguars, respectively. Both animals presented a proliferative activity varying between 1.20 and 1.30. All tissues could promote cellular detachment, reaching subconfluence in 10–15 days. This kind of information from histomorphometrical features and cell cultures can be essential for a more targeted application of ear skin cryopreservation in this species, as such information will enable understanding the action of substances on tissues during the conservation process.     

Optimizing ex situ genetic resource collections for European livestock conservation

Ex situ collections offer the potential to reduce extinction risks, affording option to society in maintaining future breeding opportunities for productivity and heritage traits. However, how much should we be seeking to collect and conserve in gene banks, and where? We developed a mathematical model to optimize logistical decisions of breed conservation choices and to evaluate alternative scenarios for efficiently re‐allocating genetic materials currently stored in different European gene banks, allowing for cross‐country collections, cost and cryogenic capacity differentials. We show how alternative allocations for the breeds that are currently stored in 11 European gene banks could reduce overall conservation costs by around 20% by selecting cryogenic banks that have relatively lower combination of fixed and collection costs, and are geographically closer to collection regions. Our results show that centralizing collections in one gene bank would double the costs, relative to collective European collections approaches. We also calculate marginal costs of collections and show that increasing diversity within the gene banks implies in higher costs per conserved breed.     

Progesterone production in mares and echographic evaluation of the corpora lutea formed after follicular aspiration

Ultrasound‐guided follicular aspiration was performed in 26 Criollo crossbred mares, followed by the evaluation of ultrasonographic images of the Corpus luteum (CL) that was formed after puncture of follicles of different diameters (Group 25–29 mm; Group 30–35 mm and Group >35 mm). Serum progesterone (P₄) concentrations were measured to determine CL function. The size of the CL was measured and the CL was classified based on the following echoscore: 1– anechoic tissue; 2– poorly defined luteal structure with low echogenicity; 3– echogenicity analogous to a luteal structure. The proportion of aspirated follicles that formed a functional CL (based on P₄ concentration) 8 days after aspiration was 57.1% (4/7; CL size 25–29 mm), 75.0% (6/8; CL size 30–35 mm) and 72.7% (8/11; CL size >35 mm), respectively (p > 0.05). The echographic scores of aspirated follicles (indicating the presence or absence of a CL) were consistent with serum P₄ concentrations (p < 0.0001). Of 26 aspirations, 18 resulted in luteal function confirmed by increased progesterone concentrations ([P₄] > 1.0 ng/ml); 17 of these mares (94.4%) had an echoscore (2–3) compatible with luteinization (p = 0.0372). Eight days after aspiration, serum [P₄] > 2.0 ng/ml was associated with high (p = 0.0056) CL echoscore (3) in 15 of 17 mares (88.2%). The echoscore used in this study was valuable as a screening test to detect the presence of a functional CL after aspiration. An echoscore of 3 served as a practical and efficient method to confirm luteinization.     

Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological findings.     

The efficiency of energy retention of broiler chickens and turkeys fed on diets with different lysine concentrations

1.?An experiment determined the effect of 6 different lysine concentrations (40, 52, 58, 65, 90 and 130 g/kg CP) in two protein concentrations (210 and 260 g/kg) on the efficiency of energy utilisation of broiler chickens and turkeys. Ninety-six male Ross 308 broiler chicks and 96 male BUT Big 6 turkey poults were fed at 90% of ad libitum during a 12 d bioassay period. At the end of the bioassay, birds were slaughtered and carcase crude protein, fat and gross energy were measured and nutrient metabolisability determined.

2.?Broiler chickens had lower AME and AME_N than turkeys. AME was higher in the 260 g/kg than in the 210 g/kg protein diet; however, no difference was evident in AME_N between any of the diets in both species. A significant species × lysine interaction indicated that the decrease in AME and AME_N with increasing lysine was greater in turkeys than in broiler chickens.

3.?Turkeys had lower weight gains and efficiencies of energy retention than broiler chickens. The effects of different lysine concentrations on growth and the efficiency of energy retention were significant and growth responses were quadratic but there were negative linear responses in the efficiency of energy retention in both species at both crude protein concentrations. The efficiency of energy retention was increased with decreasing lysine concentrations throughout the wide range of concentrations used in the experiment.     

Estimation of genetic parameters for mature weight in Angus cattle

The aim of this study was to estimate genetic parameters for BW of Angus cattle up to 5 yr of age and to discuss options for including mature weight (MW) in their genetic evaluation. Data were obtained from the American Angus Association. Only records from herds with at least 500 animals and with >10% of animals with BW at ≥ 2 yr of age were considered. Traits were weaning weight (WW, n = 81,525), yearling weight (YW, n = 62,721), and BW measured from 2 to 5 yr of age (MW2, n = 15,927; MW3, n = 12,404; MW4, n = 9,805; MW5, n = 7,546). Genetic parameters were estimated using an AIREML algorithm with a multiple-trait animal model. Fixed effects were contemporary group and departure of the actual age from standard age (205, 365, 730, 1,095, 1,460, and 1,825 d of age for WW, YW, MW2, MW3, MW4, and MW5, respectively). Random effects were animal direct additive genetic, maternal additive genetic, maternal permanent environment, and residual. Estimates of direct genetic variances (kg(2)) were 298 ± 71.8, 563 ± 15.1, 925 ± 52.1, 1,221 ± 65.8, 1,406 ± 80.4, and 1,402 ± 66.9; maternal genetic variances were 167 ± 4.8, 153 ± 6.1, 123 ± 9.1, 136 ± 12.25, 167 ± 18.0, and 110 ± 14.0; maternal permanent environment variances were 124 ± 2.9, 120 ± 4.3, 61 ± 7.5, 69 ± 11.9, 103 ± 15.9, and 134 ± 35.2; and residual variances were 258 ± 3.8, 608 ± 8.6, 829 ± 34.2, 1,016 ± 38.8, 1,017 ± 52.1, and 1,202 ± 63.22 for WW, YW, MW2, MW3, MW4, and MW5, respectively. The direct genetic correlation between WW and YW was 0.84 ± 0.14 and between WW and MW ranged from 0.66 ± 0.06 (WW and MW4) to 0.72 ± 0.11 (WW and MW2). Direct genetic correlations ranged from 0.77 ± 0.08 (YW and MW5) to 0.85 ± 0.07 (YW and MW2) between YW and MW, and they were ≥ 0.95 among MW2, MW3, MW4, and MW5. Maternal genetic correlations between WW and YW and MW ranged from 0.52 ± 0.05 (WW and MW4) to 0.95 ± 0.07 (WW and YW), and among MW they ranged from 0.54 ± 0.14 (MW4 and MW5) to 0.94 ± 0.07 (MW2 and MW3). Genetic correlations suggest that a genetic evaluation for MW may be MW2-based and that including BW from older ages could be accomplished by adjusting records to the scale of MW2.