Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis

Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.     

Estrus induction in anestrous mixed-breed goats using the “female-to-female effect”

A trial was conducted during the anestrous period in female goats to determine: (a) whether estrus can be induced in anestrous goats by administration of equine chorionic gonadotropic hormone (eCG) and PGF_2α under pen conditions and (b) whether these sexually active female goats can elicit sexual arousal in sexually inactive bucks. One hundred and fifteen pluriparous, nonlactating mixed-breed female goats were randomly assigned to one of four treatment groups: (1) administration of a single dose of 240 IU of eCG, 50 μg PGF_2α i.m., and 25 mg progesterone (P₄) (eCG; n?=?30); (2) administration of P₄ and exposure to female goats treated with eCG–PGF_2α (P₄; n?=?39); (3) administration of 0.5 ml saline and P₄ (Sal; n?=?23); and (4) P₄ plus exposure to female goats treated with saline (Con; n?=?23). After hormone administration, all goats were put together with adult sexually inactive bucks for 15 days. The percentage of goats in estrus during these 15 days was similar in eCG-treated animals and untreated animals exposed to the eCG animals (97 and 95 %). Pregnancy rate was also similar (63 vs. 64 %) between these two groups. eCG-treated goats exhibited estrus earlier (P?<?0.05) than the treated goats in contact with the eCG goats. Furthermore, eCG-treated goats had larger litters (1.9?±?0.2 vs. 1.6?±?0.1, P?<?0.05) than the untreated goats in contact with the eCG goats. These results show that fertile estrus can be induced in anestrous female goats by exposing them to female goats induced to estrus with eCG. This female–female interaction triggers the stimulation cycle leading to the sexual arousal of bucks.     

Mucosal and systemic T cell response in mice intragastrically infected with Neospora caninum tachyzoites

The murine model has been widely used to study the host immune response to Neospora caninum. However, in most studies, the intraperitoneal route was preferentially used to establish infection. Here, C57BL/6 mice were infected with N. caninum tachyzoites by the intragastric route, as it more closely resembles the natural route of infection through the gastrointestinal tract. The elicited T-cell mediated immune response was evaluated in the intestinal epithelium and mesenteric lymph nodes (MLN). Early upon the parasitic challenge, IL-12 production by conventional and plasmacytoid dendritic cells was increased in MLN. Accordingly, increased proportions and numbers of TCRαβ⁺CD8⁺IFN-γ⁺ lymphocytes were detected, not only in the intestinal epithelium and MLN, but also in the spleen of the infected mice. In this organ, IFN-γ-producing TCRαβ⁺CD4⁺ T cells were also found to increase in the infected mice, however later than CD8⁺ T cells. Interestingly, splenic and MLN CD4⁺CD25⁺ T cells sorted from infected mice presented a suppressive activity on in vitro T cell proliferation and cytokine production above that of control counterparts. These results altogether indicate that, by producing IFN-γ, TCRαβ⁺CD8⁺ cells contribute for local and systemic host protection in the earliest days upon infection established through the gastrointestinal tract. Nevertheless, they also provide substantial evidence for a parasite-driven reinforcement of T regulatory cell function which may contribute for parasite persistence in the host and might represent an additional barrier to overcome towards effective vaccination.     

Antibiotic sensitivity and biochemical characterization of Fusobacterium spp. and Arcanobacterium pyogenes isolated from farmed white-tailed deer (Odocoileus virginianus) with necrobacillosis.

Bacterial cultures from 32 living and dead farmed white-tailed deer (Odocoileus virginianus) with necrobacillosis yielded Fusobacterium necrophorum from nine individuals, F. varium from six individuals, and Arcanobacterium pyogenes from 16 individuals. The isolates were characterized biochemically using automated identification systems. Gram-stained smears suggested the presence of Fusobacterium spp. in eight cases from which organisms were not cultured. Minimum inhibitory concentration determinations in 23 strains of gram-negative anaerobic bacteria detected resistance to enrofloxacin and clindamycin. Enrofloxacin resistance was detected in A. pyogenes isolates, and although biochemical profiling indicated that the deer strains of A. pyogenes could be grouped, it is uncertain whether these biochemical characteristics correlate with antigenic or virulence factors. Deer-specific or autogenous vaccines may provide a useful alternative to generic vaccines.     

Screening genetic diseases prevalence in Braunvieh cattle

Tropical Animal Health and Production - Heritable abnormalities can cause a reduction in productive performance, structural defects, or death of the animal. There are reports of hereditary...     

Methods of collection,extender type,and freezability of semen collected from creole bulls raised in the tropical highlands of Ecuador

This study was conducted to determine the best combination between two collection method and two extenders in the cryopreservation of semen from creole bulls adapted to highlands of the Ecuadorian Andes. Sixty ejaculates from three adult Creole bulls were evaluated after collection by artificial vagina (AV) and electroejaculation (EE). Semen samples were split into two aliquots and diluted with a soy lecithin extender (Andromed®; A) or an egg yolk-containing extender (Triladyl®; T) and packed in straws of 0.25 ml with 20 × 10⁶ sperms. Optical microscopy and computer-assisted semen analysis system (CASA) were used to evaluate semen quality characteristics. The effects of collection methods and extender type as well as its interaction were evaluated by a factorial ANOVA and Bonferroni’s test. Semen samples collected with EE and frozen with T (EE-T) and A (EE-A) had greater proportion of spermatozoa with optical assessed individual progressive motility (IPM), plasmatic membrane intact (HosT), and lower tail abnormalities than those obtained with AV and frozen with the same extenders (AV-T and AV-A); however, differences were significant only between EE-A and AV-T. CASA assessment indicated that the total mobility (TM) was greater (P < 0.05) in semen samples diluted with T, although these samples had a greater proportion (P < 0.05) of sperms with local motility (LM) and fewer immobile sperms (IS), than those extended with A. Generally, semen samples obtained with EE or AV and diluted with T seems to be the best option to ciopreserve gametes of Creole bulls raised in highlands of Ecuadorian Andes.

     

Seroprevalence of spotted fever group rickettsiae in canines along the United States–Mexico border

Portions of northern Mexico are experiencing a re‐emergence of Rocky Mountain spotted fever (RMSF), a tickborne disease caused by Rickettsia rickettsii, a member of the spotted fever group of rickettsiae (SFGR). Infection with R. rickettsii can result in serious and life‐threatening illness in people and dogs. Canine seroprevalence has been used as a sentinel for human RMSF in previous studies. This study aims to quantify SFGR seroprevalence in canines in three northern Mexican states and identify risk factors associated with seropositivity. A total of 1,136 serum samples and 942 ticks were obtained from dogs participating in government sterilization campaigns and from animal control facilities in 14 Mexican cities in three states. SFGR antibodies were detected using indirect immunofluorescence antibody assays at titre values ≥1/64. Six per cent (69 dogs) showed antibodies to SFGR, with the highest seroprevalence reported in Baja California (12%), Coahuila (4%) and Sonora (4%). Dogs from Baja California had three times higher odds of having SFGR antibodies compared to dogs from Sonora (OR = 3.38, 95% CI, 1.81–6.37). Roughly one quarter (25%) of surveyed dogs were parasitized by ticks (Rhipicephalus sanguineus sensu lato) at the time of sample collection. A portion of collected ticks were tested for rickettsial DNA using polymerase chain reaction. Positive samples were then sequenced, showing evidence of SFGR including R. massiliae, R. parkeri and R. rickettsii. Dogs that spent the majority of time on the street, such as free‐roaming or community‐owned dogs, showed a greater risk of tick infestation, seropositivity, bearing seropositive ticks, and may play a pivotal role in the spread of SFGR among communities. Estimating the seroprevalence of SFGR in the canine population can help public health campaigns target high‐risk communities for interventions to reduce human RMSF cases.     

Signatures of selection in Charolais beef cattle identified by genome‐wide analysis

Charolais cattle are one of the most important breeds for meat production worldwide; in México, its selection is mainly made by live weight traits. One strategy for mapping important genomic regions that might influence productive traits is the identification of signatures of selection. This type of genomic features contains loci with extended linkage disequilibrium (LD) and homozygosity patterns that are commonly associated with sites of quantitative trait locus (QTL). Therefore, the objective of this study was to identify the signatures of selection in Charolais cattle genotyped with the GeneSeek Genomic Profiler Bovine HD panel consisting of 77 K single nucleotide polymorphisms (SNPs). A total 61,311 SNPs and 819 samples were used for the analysis. Identification of signatures of selection was carried out using the integrated haplotype score (iHS) methodology implemented in the rehh R package. The top ten SNPs with the highest piHS values were located on BTA 4, 5, 6 and 14. By identifying markers in LD with top ten SNPs, the candidate regions defined were mapped to 52.8–59.3 Mb on BTA 4; 67.5–69.3 on BTA 5; 39.5–41.0 Mb on BTA 6; and 26.4–29.6 Mb on BTA 14. The comparison of these candidate regions with the bovine QTLdb effectively confirmed the association (p < 0.05) with QTL related to growth traits and other important productive traits. The genomic regions identified in this study indicated selection for growth traits on the Charolais population via the conservation of haplotypes on various chromosomes. These genomic regions and their associated genes could serve as the basis for haplotype association studies and for the identification of causal genes related to growth traits.     

Induction of CIRBP expression by cold shock on bovine cumulus–oocyte complexes

The aim of this study was to induce the cold‐inducible RNA‐binding protein (CIRBP) expression on cumulus–oocyte complexes (COCs) through exposure to a sub‐lethal cold shock and determine the effects of hypothermic temperatures during the in vitro maturation of bovine oocytes. Nuclear maturation, cortical granule redistribution and identification of cold‐inducible RNA‐binding protein (CIRBP) were assessed after 24 hr of in vitro maturation of control (38.5°C) and cold‐stressed oocytes (33.5°C). The presence of CIRBP was assessed by Western blot in COCs or denuded oocytes and their respective cumulus cells. Based on the odds ratio, cold‐stressed oocytes presented higher abnormal cytoplasmic distribution of cortical granules and nuclear maturation than the control group. Although CIRBP was detected in both control and cold‐stressed groups, cold‐stressed COCs had 2.17 times more expression of CIRBP than control COCs. However, when denuded oocytes and cumulus cells were assessed separately, CIRBP only was detected in cumulus cells in both groups. In conclusion, cold shock induced CIRBP expression, but it negatively affected nuclear maturation and cortical granule distribution of bovine oocytes. Moreover, the expression of CIRBP was only identified in cumulus cells but not in oocytes.