Evaluation of single and comparative intradermal tuberculin tests for tuberculosis eradication in caprine flocks in Castilla y León (Spain)

Goats can act as reservoirs for tuberculosis (TB) infection. The main etiological agents of TB in goats are Mycobacterium caprae and Mycobacterium bovis and they infect also a wide range of domestic and wild animals and humans. Control programmes based mainly on the application of single and comparative intradermal tuberculin (SIT and SCIT respectively) tests are being implemented in certain regions of Spain with a high density of caprine flocks as Castilla y León, including goats with epidemiological relationship with cattle. The aim of this study was to evaluate the performance of the intradermal tests in naturally TB-infected caprine flocks from this region. The study was performed using data from 17,450 goats in 54 different flocks that were classified as TB-infected in the control programmes executed in 2010 and 2011. Data from 1237 goats from 7 dairy flocks depopulated after the first intradermal testing were used to estimate the sensitivity (Se) using bacteriology as the gold-standard. Overall Se of the SIT test using the severe interpretation was 43.9% (CI 95%, 40.4–47.4) and decreased to 38.8% (CI 95%, 35.5–42.3) using the standard interpretation. Overall Se of the SCIT test ranged between 21.3% (CI 95%, 17.6–25.4) and 7% (CI 95%, 4.9–9.8) depending of the interpretation criteria. A significant weak positive correlation was found between age and skin fold thickness (Spearman’s test p < 0.05). Results from this study yielded, in general, low Se values probably due the systematic detection and slaughter of reactors as a consequence of the eradication programme in previous years or the presence of factors that may interfere in the diagnosis. Therefore, these results suggest the necessity of including ancillary diagnostic tools and/or strict interpretation criteria to maximize detection of positive animals in infected settings.     

Evaluation of hepatitis E virus infection between different production systems of pigs in Brazil

The objectives of this study were to (1) investigate the occurrence of hepatitis E virus (HEV) in pigs from large-scale and family-scale farms, (2) genetically characterize the strains isolated, and (3) study the pathogenesis of swine HEV infection via immunohistochemistry. A total of 50 pigs from 10 farms in Mato Grosso State, Brazil were divided according to type of production system into either large-scale farms (n?=?5) or family-scale farms (n?=?5). Samples of liver, gallbladder, small and large intestines, bile, and feces from the pigs were analyzed by nested PCR with primers targeting the ORF2 region of HEV and by immunohistochemistry. Of the eight HEV-positive samples from pigs of family-scale farms, phylogenetic analysis revealed that seven of the swine HEV isolates clustered with subtype 3b of genotype 3 and one isolate was categorized with subtype 3 f. The HEV antigen was detected mainly in the small intestine samples from family-scale farms, suggesting an early stage HEV infection. HEV was not detected in the samples of pigs from large-scale farms, reinforcing the need for additional studies to evaluate the risk of transmission of HEV to humans from pigs from family-scale farms in Mato Grosso State.     

Prevalence and risk factors associated with goat gastrointestinal helminthiasis in the Sertão region of Paraíba State,Brazil

Gastrointestinal helminthiasis represents an obstacle to goat raising, causing severe damage to herds such as growth retardation, weight loss, and even death. In this study, we aimed to determine the prevalence and risk factors associated to goat gastrointestinal helminthiasis in the Sertão region of Paraíba State, Brazil. A total of 256 goats from 54 farms were systematically sampled. Blood and fecal samples were collected from each animal for egg per gram (EPG), larval culture, and packed cell volume (PCV) analyses. We found that 79.3 % of the goats investigated were parasitized with gastrointestinal helminths. Significant correlation (p?=?0.004) was observed between the EPG and PCV of the animals studied, and it was observed that the EPG increases as the PCV decreases. In the larval culture, the most prevalent helminth was Haemonchus sp. (83.2 %). Age and sex were significant variables (p?≤?0.20) for the development of gastrointestinal helminths: 86.8 % of animals over 36 months of age and 81.7 % of females were infected. The variable type of animal exploitation was also significant, with 90.3 % (p?≤?0.20) of the animals presenting double suitability (milk and meat). The Sertão region of Paraíba State presents high prevalence of gastrointestinal helminthiasis in goats, and age and type of animal exploitation are the most relevant risk factors to the development of these parasites.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

First report of Enterocytozoon bieneusi from dairy cattle in Argentina

Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.     

Neospora caninum infection as a cause of reproductive failure in a sheep flock

Neospora caninum has been detected only sporadically in cases of ovine abortion, and it has therefore traditionally been considered as an unimportant parasite in small ruminants. This study was carried out with the aim of identifying the pathogen causing serious reproductive problems on a commercial sheep farm. Sera from all rams and ewes tested negative for antibodies against Border disease virus, Schmallenberg virus and Coxiella burnetii, and infections by these agents were therefore ruled out. Nevertheless, seropositivity to N. caninum and/or Toxoplasma gondii was detected, although the seroprevalence was higher in the case of N. caninum. The percentage of lambings and the number of lambs per dam were significantly lower in ewes that were seropositive to N. caninum while no effect on these parameters was detected in ewes that were seropositive to T. gondii. There was also no evidence of infection by T. gondii in the foetal/lamb tissues analyzed by PCR and/or immunohistopathological techniques. On the contrary, the DNA of N. caninum was detected in 13 out of 14 foetuses/lambs descendant from dams seropositive to this parasite. Characteristic lesions caused by N. caninum and/or its antigen were also detected. Genotyping of the N. caninum DNA revealed only two closely related microsatellite multilocus genotypes. The results clearly demonstrate that infection by N. caninum was the cause of the low reproductive performance of this sheep flock.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Amino acid digestibility of heat damaged distillers dried grains with solubles fed to pigs

The primary objective of this experiment was to determine the effects of heat treatment on the standardized ileal digestibility(SID) of amino acids(AA) in corn distillers dried grains with solubles(DDGS) fed to growing pigs. The second objective was to develop regression equations that may be used to predict the concentration of SID AA in corn DDGS. A source of corn DDGS was divided into 4 batches that were either not autoclaved or autoclaved at130°C for 10, 20, or 30 min. Four diets containing DDGS from each of the 4 batches were formulated with DDGS being the only source of AA and CP in the diets. A N-free diet also was formulated and used to determine the basal endogenous losses of CP and AA. Ten growing pigs(initial BW: 53.5 ± 3.9 kg) were surgically equipped with a T-cannula in the distal ileum and allotted to a replicated 5 × 4 Youden square design with 5 diets and 4 periods in each square. The SID of CP decreased linearly(P 0.05) from 77.9% in non-autoclaved DDGS to 72.1, 66.1, and 68.5% in the DDGS samples that were autoclaved for 10, 20, or 30 min, respectively. The SID of lysine was quadratically reduced(P 0.05) from 66.8% in the non-autoclaved DDGS to 54.9, 55.3, and 51.9% in the DDGS autoclaved for 10, 20, or30 min, respectively. The concentrations of SID Arginine, Histidine, Leucine, Lysine, Methionine, Phenylalanine, or Threonine may be best predicted by equations that include the concentration of acid detergent insoluble N in the model(r2= 0.76, 0.68, 0.67, 0.84, 0.76, 0.73, or 0.54, respectively). The concentrations of SID Isoleucine and Valine were predicted(r2= 0.58 and 0.54, respectively) by the Lysine:CP ratio, whereas the concentration of SID Tryptophan was predicted(r2= 0.70) by the analyzed concentration of Tryptophan in DDGS. In conclusion, the SID of AA is decreased as a result of heat damage and the concentration of SID AA in heat-damaged DDGS may be predicted by regression equations developed in this experiment.     

Escherichia coli challenge and one type of smectite alter intestinal barrier of pigs

An experiment was conducted to determine how an E. coil challenge and dietary clays affect the intestinal barrier of pigs. Two groups of 32 pigs （initial BW： 6.9 ± 1.0 kg） were distributed in a 2 x 4 factorial arrangement of a randomized complete block design （2 challenge treatments： sham or E. coil, and 4 dietary treatments： control, 0.3% smectite A, 0.3% smectite B and 0.3% zeolite）, with 8 replicates total. Diarrhea score, growth performance, goblet cell size and number, bacterial translocation from intestinal lumen to lymph nodes, intestinal morphology, and relative amounts of sulfo and sialo mucins were measured. The E. coli challenge reduced performance, increased goblet cell size and number in the ileum, increased bacterial translocation from the intestinal lumen to the lymph nodes, and increased ileal crypt depth. One of the clays （smectite A） tended to increase goblet cell size in ileum, which may indicate enhanced protection. In conclusion, E. coli infection degrades intestinal barrier integrity but smectite A may enhance it.     

Morphological and biochemical changes in the muscle of the marine shrimp Litopenaeus vannamei during the molt cycle

Morphological changes and biochemical composition of abdominal muscles over the molt cycle were investigated in juvenile Litopenaeus vannamei. Eight molt stages were characterized and clear uropod images are presented. Molt frequency was highly correlated with the age of the shrimp. Juvenile shrimps appeared to molt faster at one month of age (4.6 ± 0.5 days/cycle), slow to 11.8 ± 1.7 days/cycle at three months of age, and reach a long molt cycle at six months (17.2 ± 2.7 days/cycle). Myofiber cross-sectional images revealed specific morphological changes in abdominal muscle associated with each molt stage. Expanded fiber size was observed during intermolt stages. Water content and total soluble proteins were balanced throughout the molt cycle. Total DNA concentration increased in intermolt and premolt, while total RNA concentration remained stable except in late-premolt stages. SDS-PAGE analysis showed high levels of actin and myosin in postmolt, reaching a plateau in intermolt and declining in premolt. These results suggest the occurrence of muscle fiber rearrangement in both the premolt and postmolt stages. Abdominal muscle buildup occurs mostly during the intermolt stage.