Post mortem parasitological evaluation of dogs seroreactive for Leishmania from Rio de Janeiro, Brazil

A parasitological study was conducted on 66 dogs seroreactive for Leishmania captured as a control measure of visceral leishmaniasis in the State of Rio de Janeiro, Brazil. Biological samples from different anatomical sites were collected during autopsy of the animals and cultured on biphasic medium (NNN/Schneider). The Leishmania isolates were characterized by isoenzyme electrophoresis. Leishmania was isolated from 80.3% of the animals: 12 animals with Leishmania (Viannia) braziliensis isolated exclusively from cutaneous lesions, 39 with L. (L.) chagasi isolated from different sites in the same animal, and 2 with simultaneous isolation of L. (V.) braziliensis from cutaneous lesions and L. (L.) chagasi from different sites. Isolation in culture revealed the absence of Leishmania parasites in 13 animals. The results obtained confirm the existence of mixed infections in dogs in Rio de Janeiro and indicate the need to complement the investigation of seroreactive dogs using methods for the parasitological diagnosis and identification of Leishmania species.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.     

Genomewide association for a dominant pigmentation gene in sheep

Most published genomewide association studies (GWAS) in sheep have investigated recessively inherited monogenic traits. The objective here was to assess the feasibility of performing GWAS for a dominant trait for which the genetic basis was already known. A total of 42 Manchega and Rasa Aragonesa sheep that segregate solid black or white coat pigmentation were genotyped using the SNP50 BeadChip. Previous analysis in Manchegas demonstrated a complete association between the pigmentation trait and alleles of the MC1R gene, setting an a priori expectation for GWAS. Multiple methods were used to identify and quantify the strength of population substructure between black and white animals, before allelic association testing was performed for 49 034 SNPs. Following correction for substructure, GWAS identified the most strongly associated SNP (s26449) was also the closest to the MC1R gene. The finding was strongly supported by the permutation tree‐based random forest (RF) analysis. Importantly, GWAS identified unlinked SNP with only slightly lower p‐values than for s26449. Random forest analysis indicated these were false positives, suggesting interpretation based on both approaches was beneficial. The results indicate that a combined analytical approach can be successful in studies where a modest number of animals are available and substantial population stratification exists.     

Prevalence and antimicrobial resistance of MRSA across different pig age groups in an intensive pig production system in Australia

This observational study aimed to determine MRSA prevalence using strain‐specific real‐time PCR at the pig level, stratified by age groupings, within a pig enterprise. A total of 658 samples were collected from individual pigs (n = 618) and the piggery environment (n = 40), distributed amongst five different pig age groups. Presumptive MRSA isolates were confirmed by the presence of mecA, and MALDI‐TOF was performed for species verification. All isolates were tested against 18 different antimicrobials. MRSA was isolated from 75.2% (95% CI 71.8–78.6) of samples collected from pigs, and 71% of the MRSA isolates from this source were identified as community‐associated (CA)‐MRSA ST93, while the remainder were livestock‐associated (LA)‐MRSA ST398. Amongst environmental isolates, 80% (CI 64.3–95.7) were ST93 and the remainder ST398. All MRSA isolates from pigs and the environment were susceptible to ciprofloxacin, gentamicin, linezolid, mupirocin, rifampicin, sulfamethoxazole–trimethoprim, teicoplanin and vancomycin. Phenotypic rates of resistance were penicillin (100%), clindamycin (97.6%), erythromycin (96.3%), ceftiofur (93.7%), chloramphenicol (81.2%), tetracycline (63.1%) and amoxicillin–clavulanate (63.9%). A low prevalence of resistance (9.2%) was observed against neomycin and quinupristin–dalfopristin. The probability of MRSA carriage in dry sows (42.2%) was found to be significantly lower (p < .001) when compared to other age groups: farrowing sows (76.8%, RR1.82), weaners (97.8%, RR 2.32), growers (94.2%, RR 2.23) and finishers (98.3%, RR 2.33). Amongst different production age groups, a significant difference was also found in antimicrobial resistance for amoxicillin–clavulanate, neomycin, chloramphenicol and tetracycline. Using the RT‐PCR assay adopted in this study, filtering of highly prevalent ST93 and non‐ST93 isolates was performed at high throughput and low cost. In conclusion, this study found that weaner pigs presented a higher risk for CA‐MRSA and antimicrobial resistance compared to other age groups. These findings have major implications for how investigations of MRSA outbreaks should be approached under the One‐Health context.     

Pharmacokinetics of cefquinome in tilapia (Oreochromis niloticus) after a single intramuscular or intraperitoneal administration

The pharmacokinetics of cefquinome was studied in plasma after a single dose (10 mg/kg) of intramuscular (i.m.) or intraperitoneal (i.p.) administration to tilapia (Oreochromis niloticus) in freshwater at 30 °C. Ten fish per sampling point were examined after treatment. The data were fitted to two‐compartment open models following both routes of administration. The estimates of total body clearance (CL/F), volume of distribution (Vd/F), and absorption half‐life (T_1/2ka) were 0.049 and 0.037 L/h/kg, 0.41 and 0.33 L/kg, and 0.028 and 0.035 h following i.m. and i.p. administration, respectively. After i.m. injection, the elimination half‐life (T_1?2β) was calculated to be 5.81 h, the maximum plasma concentration (C_max) to be 49.40 μg/mL, the time to peak plasma cefquinome concentration (T_max) to be 0.14 h, and the area under the plasma concentration–time curve (AUC) to be 204.6 μg h/mL. Following i.p. administration, the corresponding estimates were 6.05 h, 44.39 μg/mL, 0.17 h and 267.8 μg h/mL. The minimum inhibitory concentrations of cefquinome, determined for 30 strains of Streptococcus agalactiae isolated from diseased tilapia, ranged from 0.015 to 0.12 μg/mL. Results from these studies support that 10 mg cefquinome/kg body weight daily could be expected to control tilapia bacterial pathogens inhibited in vitro by a minimal inhibitory concentration value of ≤2 μg/mL.     

Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.