Effect of Mucine 4 and Fucosyltransferase 1 genetic variants on gut homoeostasis of growing healthy pigs

Putative genetic markers have been associated with ETEC F4 (Mucine 4 [MUC4]; MUC4^GG;CG as susceptible; MUC4^CC as resistant) and F18 (Fucosyltransferase 1 [FUT1]; FUT1^GG;AG as susceptible; FUT1^AA as resistant) resistances respectively. In this study, 71 post‐weaning pigs were followed from d0 (35 days old) to d42 (77 days of age) to investigate the effect of MUC4 or FUT1 genotypes on the mid‐jejunal microbiota composition, pigs expression of genes related to inflammation (IL8, GPX2, REG3G, TFF3, CCL20 and LBPI) and glycomic binding pattern profile (Ulex europaeus agglutinin I [UEA] fucose‐binding lectin and peanut agglutinin [PNA] galactose‐specific), and on blood plasma targeted metabolomics profile, faecal score and performance parameters of growing healthy pigs. The MUC4 and FUT1 resistant genotypes improved the pigs’ growth performance and had firmed faecal score susceptible genotypes in d0–d21 period. Pigs with MUC4^GG genotype had a higher jejunal expression of genes relate to immune function (CCL20 and REG3G) than MUC4^CG and MUC4^CC pigs (p < 0.05). MUC4^CG pigs had higher expression of TFF3 (implicated in mucosal integrity) than MUC4^GG and MUC4^CC (p < 0.05). FUT1 influenced the alpha‐ and beta‐jejunal microbial indices. The FUT1^AA group had a higher number of operational taxonomic units (OTUs) belonging to Lactobacillus genus, while FUT1^GG group had a higher number of OTUs belonging to Veillonella genus. MUC4^CC pigs had lower scores for UEA on brush borders and goblet cells in villi than MUC4^GG (p < 0.05). FUT1^AA pigs had lower UEA positivity and higher PNA positivity on brush borders and goblet cells than FUT1^AG and FUT1^GG (p < 0.05). Both FUT1 and MUC4 influenced the metabolic profile of healthy pigs. Results highlight the role of MUC4 and FUT1 on pig intestinal homoeostasis and improved the knowledge regarding the potential interaction between host genetics, gut microbiota composition and host metabolism in a healthy status.     

Retrospective Biomolecular Investigation of Coxiella burnetii and Leptospira spp. DNA in Cases of Abortion,Stillbirth and Neonatal Mortality in Dogs and Cats

Abortion and neonatal mortality are events that can occur in breeding bitches and queens. It has been reported that up to 55% and 33% of these cases remain without a known cause, respectively, in canine and feline pregnancies. Unusual abortigenic and potentially zoonotic agents, including Coxiella burnetii and Leptospira spp., may be involved in these cases. C. burnetii is able to cause reproductive disorders in cattle, sheep and goats, and cases of abortion have been observed in dogs and cats. Moreover, several outbreaks of C. burnetii infection in humans have been caused by delivering bitches and queens, and some of these animals experienced abortion. Leptospira interrogans sensu lato is able to cause abortion or stillbirth in several animal species and its abortigenic role has occasionally been described in bitches and queens. The aim of this study was to search for C. burnetii and Leptospira spp. DNA in a retrospective series of 103 cases of canine and feline abortion, stillbirth, and neonatal mortality submitted for the identification of possible infectious agents. One hundred and fifty-one specimens were tested using PCR assays and found negative for C. burnetii and Leptospira DNA. However, in 49 samples (47.6%) other infectious causes of abortion, stillbirth, and neonatal mortality were identified. These results showed that C. burnetii and Leptospira spp. are probably not common abortigenic agents or causes of neonatal deaths in dogs. However, given the potential abortigentic and zoonotic role of these agents, surveillance of canine and feline abortion, stillbirth, and neonatal mortality could be advisable for a systematic investigation of these events.     

Timely adjuvant chemotherapy improves outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy

Timely delivery of adjuvant chemotherapy has been shown to be advantageous in many human cancers and canine osteosarcoma. Adjuvant chemotherapy has been shown to improve outcome for canine splenic hemangiosarcoma. The aim of this retrospective study was to investigate whether timely adjuvant chemotherapy administration resulted in better outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy. Medical records were searched for dogs with non-metastatic, splenic hemangiosarcoma that received splenectomy and adjuvant chemotherapy. The number of days from surgery to the first chemotherapy dose (StoC) was evaluated to identify the cut-off value associated with the best survival advantage. StoC and other possible prognostic factors were tested for influence on time to metastasis (TTM) and overall survival (OS). Seventy dogs were included. Median StoC was 20 days (range: 4–70). The time interval associated with the greatest survival benefit was 21 days. Median TTM and OS of dogs with StoC ≤ 21 days were significantly longer than those with StoC >21 days (TTM: 163 vs. 118 days, p = .001; OS: 238 vs. 146 days, p < .001). On multivariable analysis, StoC >21 days was the only variable significantly associated with increased risk of tumour progression (HR 2.1, p = .010) and death (HR 2.3; p = .008). Starting adjuvant chemotherapy within 21 days of surgery may be associated with a survival benefit in dogs with non-metastatic splenic hemangiosarcoma, possibly due to the early targeting of newly recruited metastatic cells after surgery.     

A simple molecular method for discriminating common filarial nematodes of dogs (Canis familiaris)

Accurate diagnosis of canine filariosis is essential for choosing correct therapeutic approach. Therefore, reliable methods for discriminating among the different filarial infections in dogs are needed. The authors report simple and highly specific molecular methods that identify the three most common filarial nematodes of European dogs: Dirofilaria immitis, D. repens and Acanthocheilonema (syn. Dipetalonema) reconditum, based on (1) PCR amplifications of mitochondrial DNA (12S rDNA and coxI) with general filarial primers followed by digestion with restriction enzymes that generates band polymorphisms clearly discriminating the three species and (2) PCR amplifications with species-specific primers to support the restriction analysis, in particular in the case of multiple infections.     

Three cases of immune-mediated adnexal skin disease treated with cyclosporin

Cyclosporin is currently considered a new and interesting drug in veterinary dermatology for the treatment of immune-mediated skin diseases, and a safe and effective alternative to immunosuppressive therapy with glucocorticoids. The authors report a case of granulomatous folliculitis and furunculosis and of sebaceous adenitis in two cats and a case of alopecia areata in a dog, successfully controlled with cyclosporin.     

Use of capecitabine after renal allograft transplantation in dog erythrocyte antigen-matched dogs

OBJECTIVES: To investigate the use of a capecitabine (CAP)-based regimen after renal transplantation in dogs. STUDY DESIGN: Prospective, pilot study. ANIMALS: Healthy, unrelated, dog erythrocyte antigen (DEA)-matched, adult beagles. METHOD: Standard heterotopic renal transplantation with native nephrectomy was performed in 7 dogs. Dogs received oral, twice daily, CAP (250 mg/m2), cyclosporine-A (CsA) (4 mg/kg), ketoconazole (5 mg/kg), and prednisolone (0.25 mg/kg). After 90 days the surviving dogs were euthanatized and complete necropsy was performed. RESULTS: Seven transplants were performed. All dogs survived surgery. Six dogs had acute neurotoxicity, which resulted in death or euthanasia of 2 dogs within 2 days of surgery. In the remaining dogs, toxicity resolved rapidly with cessation of drug administration. Thereafter, modification of the regimen minimized toxicity. The 5 remaining dogs survived to study end; 4 dogs had no evidence of graft rejection. Necropsy examination was mostly unremarkable in all dogs. There were no major changes in CBC or biochemical values, except for a significant increase in serum calcium. CONCLUSIONS: CAP appeared well tolerated in most dogs. Toxicity occurred but abated with modification of the drug regimen. Efficacy for postoperative immunosuppression cannot be determined by this study, although results are promising. CLINICAL RELEVANCE: CAP-CsA-prednisolone is an effective, oral immunosuppressive regimen for prevention of acute allograft rejection in DEA-matched beagles. Further studies on dose, toxicity, and efficacy compared with current immunosuppressive regimens are needed before use in clinical practice.     

Comparison of 4 fixation and staining methods for the cytologic evaluation of ear canals with clinical evidence of ceruminous otitis externa

BACKGROUND: Swab cytology of ear canals is one of the most useful and rapid methods to assess the presence of external ear infections. Smears are generally stained with rapid Romanowsky-type stains, with or without prior heat fixation. OBJECTIVES: The aim of this study was to compare 4 different methods of fixation and staining of ear swab cytology samples from dogs. METHODS: Eight dogs with otitis externa were selected from a dermatology referral population. A cotton swab was used to obtain ceruminous material from 12 ear canals. Four smears of each swab were prepared on glass slides (randomly identified as A, B, C, or D) and air-dried for cytologic examination. Samples marked A were stained with Dip Quick (Jorgensen Laboratories Inc, Loveland, CO, USA) after heat fixation; samples marked B were stained without heat fixation; samples marked C were heat-fixed and dipped only in the counterstain (the blue reagent) of Dip Quick; and samples marked D were dipped only in the counterstain, without heat fixation. Ten high-power fields (hpf; X100 oil immersion objective) in each slide were evaluated by 2 observers, and total numbers of keratinocytes, yeast, bacteria, and neutrophils were counted. Statistical comparison was performed using an ANOVA model applied after verifying the normal distribution of the data, and using nonparametric sign tests and Wilcoxon signed rank tests. RESULTS: No statistically significant differences were observed in the numbers of keratinocytes, yeast, bacteria, or neutrophils among the 4 staining methods (P > .05), although significant interobserver differences were found. CONCLUSION: We conclude that heat fixation does not improve the quality of ceruminous ear swab samples for cytologic evaluation, and propose a 1-step dip in the blue reagent alone as a rapid method of staining samples from canine ear canals.