Incidental Detection of Onchocerca Microfilariae in Donkeys (Equus asinus) in Italy: Report of Four Cases

This paper reports the occurrence for the first time in Italy of autochthonous Onchocerca infection in donkeys. Four jennies, bred on the same farm, were referred to the Veterinary Teaching Hospital of Pisa for a check-up on ovarian activity (n = 3) or for veterinary support during the delivery (n = 1). Microfilariae were incidentally detected during the blood smear examination of one jenny. Peripheral blood samples were then collected from the other three jennies and the presence of microfilariae was investigated by Knott's test. Circulating unsheathed microfilariae were identified in all the animals. The level of microfilaraemia was between 1 and 31 microfilariae in 2 mL of blood. Hematological changes showed moderate eosinophilia in one case or both remarkable eosinophilia and basophilia in another case. Based on molecular findings by PCR and sequencing, the microfilariae showed 98% sequence similarity with Onchocerca sp. in the NCBI GenBank database (Accession No.: MK541848.1). The present report provides evidence that Onchocerca is an etiological agent of parasitic infection in donkeys in Italy. Our findings highlight the importance of screening in donkeys for Onchocerca even in the absence of clinical indications

Prevalence of Dog Erythrocyte Antigen 1 in 7, 414 Dogs in Italy

The study aim was to establish the prevalence of DEA 1, the most immunogenic and clinically important blood group in canine blood transfusion, in 7,414 dogs from Italy. The potential sensitization risk following a first transfusion and the acute reaction risk following a second transfusion given without a cross-matching and blood typing test were also calculated. Dogs tested were purebred (4,798) and mongrel (2,616); 38.8% were DEA 1 negative and 61.2% were DEA 1 positive. High prevalence for DEA 1 positive blood type was found in Ariegeois and English Setter, whereas German Shepherd and Boxer had higher DEA 1 negative blood type. Breeds with blood type never reported before included French Brittany Spaniel and Pug showing a high prevalence of DEA 1 positive type, while French Bulldog and West Highland White Terrier were more often DEA 1 negative. Just 48.8% of purebred and 13.9% of mongrel dogs were considered as prospective blood donors based upon their blood type.Most of the breeds had a sensitization risk of 20.0–25.0%. Rottweiler and Ariegeois had less risk of sensitization (9.4 and 4.2%) and the minor risk of an acute transfusional reaction (0.9–0.2%).The prevalence of DEA 1 positive and negative dogs in Italy agrees withmost of the data already reported in the literature

SUCCESSFUL MANAGEMENT OF ACUTE BABESIOSIS IN A DOG

Canine babesiosis is a tick-borne disease caused by Babesia spp. Dogs with uncomplicated babesiosis typically show pale mucous membranes, fever, anorexia, depression, water-hammer pulse, and splenomegaly. The complicated form can include acute renal failure, cerebral babesiosis, coagulopathy, icterus and hep- atopathy, immune-mediated hemolytic anemia (IMHA), acute respiratory distress syndrome (ARDS), hemo- concentration. This case report describes the presentation, diagnosis, and management of acute systemic inflammatory response syndrome (SIRS) in a dog affected by Babesia canis. A Border Collie, intact male, 8-years-old, was presented in emergency setting showing weakness, anorexia and ’pigmenturia’ started 2 days before. The dog was used as cattle dog in Piedmont region and recently moved to Tuscany. Dog showed fever (38.

INTESTINAL MICROBIOTA IN LYMPHOMA: A COMPARISON IN HEALTY DOGS AND DOGS WITH NON HODGKIN LYMPHOMA

Background and objectives Animal models play a key role in understanding the importance of gut microbiome in immune development and composition as well as to reinforce the relationship between the microbiota and health and disease. Intestinal bacteria have been implicated in several types of cancer. Regardless, microbes influence immune cells directly, indirectly, or both, and increased lymphocyte proliferation can lead to a higher chance of aberrant DNA replication. This particularly occurs with some B lymphocytes which are innately vulnerable to genetic instability and activation. Methods We analyzed the microbiome (by using 16S rRNA gene sequencing and qPCR assays) of naturally voided fecal samples from 12 healthy and 12 Non Hodgkin Lymphoma (NHL) dogs in order to evaluate the microbiota composition using a dysbiosis index. An index value greater than 2 indicates dysbiosis, while below 0 indicates normal microbiota. Results. Significant differences were observed when comparing the fecal microbiota structure of all healthy dogs vs NHL dogs (ANOSIM; P<0.05). Specifically, differences were observed for Faecalibacterium (P<0.001) with concentrations higher in healthy vs NHL dogs. The dysbiosis index was significantly lower (p=0.007) in healthy vs NHL dogs (mean, SD: H2.6, 2.0 vs 1.7, 3.2), respectively. Conclusion Interestingly, lower levels of Fecalibacterium prausnitzii were recently found in humans with some chronic colonic conditions as well as colorectal cancer (P < 0.001) compared with healthy subjects. This study showed that NHL have a increased dysbiosis index, indicating dysbiosis. Animal models of cancer can be critical in order to demonstrate a link between the microbiome and carcinogenesis

Fecal microbiota differences in Non-Hodgkin Lymphoma (NHL) affected dogs: preliminary results.

Animal models play an essential role in understanding the importance of gut microbiome in immune development and composition, and play a key role to reinforce the relationship between the microbiome and health and disease [3]. Non-Hodgkin Lymphoma (NHL) is the most common hematopoietic malignancy in dogs, caused by clonal proliferation of lymphocytes in solid organs [2]. Whether microbes influence immune cells directly, indirectly, or both, increased lymphocyte proliferation can lead to a higher chance of aberrant DNA replication, particularly in some B lymphocytes which are innately vulnerable to genetic instability and activation. Oxidative stress caused by intestinal microbiota, either directly or indirectly through the immune system, can also affect tumorigenesis, thus, the microbiota can affect several pathways associated with lymphomagenesis [4]. The optimal responses to cancer therapy require an intact commensal microbiota that mediates its effects, by modulating myeloid derived cell functions in the tumour microenvironment [1]. In our study design we analysed the microbiome (by using 16S rRNA gene 454-pyrosequencing and qPCR assays) of naturally voided fecal samples from 6 healthy dogs, 8 NHL dogs before and 4 NHL (of the eight) dogs after induction phase of chemotherapy (cyclophosphamide, vincristine, and prednisolone) plus probiotics (Sivoy TM). Several statistical significances were observed compared the fecal microbiome of healthy dogs vsNHL dogs before chemotherapy. In particular, differences were observed for Bifidobacteria (p=0.0001), Lactobacillus (p=0.0001), Faecalibacterium (p=0.0005), Bacteroidetes (p=0.0480), and Fusobacterium (p=0.0025), which concentrations were higher in healthy dogs compared to NHL dogs. On the contrary, the concentration of Clostridium perfrigens was greater in NHL dogs compared to healthy dogs (p=0.0326). No statistical differences for total bacteria, Escherichia coli, Blautia, and Ruminococcaceae were found. Microbiome shift (total bacteria, Bifidobacteria, Lactobacillus, Faecalibacterium, Bacteroidetes, Fusobacterium, Escherichia coli, Blautia, Ruminococcaceae, and Clostridium perfrigens) of fecal samples were also compared before and after induction phase of chemotherapy plus probiotics (Sivoy TM probiotic mix Slab51, containing 8 strains of lactic acid bacteria and bifidobacteria dosed at 200 billion per stick) but no statistical significance was found. In order to understand microbiome’s changes in NHL affected dogs treated with standard protocol plus probiotics, a larger number of stool samples before and after treatment, from a greater number of animals, should be investigated. The fact that an increased number of lymphomas are becoming associated with bacterial infections underscores the need for more studies involving microbes and lymphoma and about the use of probiotics to restore normal microbiota in affected dogs

Comparison of Standard Protocols for the Treatment of Canine Leishmaniasis in an Endemic Area with and Without Zinc Oral Supplementation

Successful treatment of canine leishmaniasis (CanL) depends on an effective cellular immune response. Zinc is an essential trace element for the immune system and in dogs with clinical leishmaniasis low serum zinc levels have been reported. The aim of this work was to evaluate the effect of zinc oral administration during treatment of CanL. Eighteen dogs from an endemic area were enrolled showing clinical signs of leishmaniasis and diagnosed by positive parasitological and serological tests. Dogs were subdivided in three treatment groups: MA, meglumineantimoniate 50 mg/kg SC for 30 days with allopurinol 10 mg/kg PO BID for 7 months; MZ, meglumineantimoniate 50 mg/kg SC BID for 30 days with zinc 2.2 mg/kg/die PO for 12 months; MAZ, same as MA group plus supplemented with zinc 2.2 mg/kg/die PO for 12 months. Each dog was monitored for 12 months using clinical and skin scores and some blood biochemical markers. Dogs in MZ and MAZ group showed a better and earlier improvement of clinical and skin scores in comparison to control dogs (MA group). Among few blood markers studied (hemoglobin, albumin, γ globulins and A/G ratio) dogs in MAZ group did improve and earlier than other groups suggesting that zinc improves the condition where allopurinol is also present. The supplementation of zinc in the treatment protocol for CanL increased the serum zinc concentrations. In addition, preliminary data showed in group MZ and MAZ dogs a faster response to therapy and the elongation of the disease-free interval time

INVESTIGATION OF CRP AND OTHER HEMATIC INFLAMMATION MARKERS IN DOGS

The systemic reaction to acute inflammation, also known as acute phase response, induces some hemato-biochemical changes, which can be evidenced in laboratory findings. The C Reactive Protein (CRP) is a main acute phase response protein elective in dogs to detect inflammatory disease. Other blood parameters have been described as useful inflammation markers i.e. Fibrinogen and Leukocytes (1, 2, 3). The aim of this retrospective study was to investigate the CRP values in comparison to Fibrinogen (Fib), Albumin (Alb), and Iron (Fe) values, total White Blood Cell (WBC), Segmented Neutrophil (NeuSeg) and Band Neutrophil (Band) counts, and the occurrence of Toxic Neutrophils (Neu TOX), Activated Monocytes (Mon ATT), and Reactive Lymphocytes (Linf REA) in blood smears. For this purpose, data of 1,837 blood samples was collected over a three-year period (2012-2015). Data collected for each sample included: Fib, Alb, Fe, WBC, NeuSeg, Band, Neu TOX, Mon ATT, Linf REA and CRP, as well as information regarding dog’s age, breed, and gender. Blood samples were divided into 2 groups: "inflammatory";; CRP ≥0.30 mg/dL (#1080) and non-inflammatory;; CRP ≤0.29 mg/dL (#757). The 2 groups were compared using: Chi squared for sex, breed, and age; Relative risk (RR) for age; Spearman Rank correlation test (SRct) for all parameters studied; Multiple regression (MR) to assess the relationship between CRP and other inflammation markers; Receiver Operating Characteristic (ROC) curves for diagnostic accuracy of each parameter in comparison to CRP (MedCalc®, 14.8). Dogs belonging to inflammatory group were significantly older (>7 years old) than those of non-inflammatory group (P<0.05), (RR, 1.38). Low yet significant (p<0.01) correlations between CRP and the other markers were noted using the SRct (R): CRP/Fib, +0.26; CRP/NeuSeg, +0.26; CRP/WBC, +0.24; CRP/Alb, -0.21; CRP/Band, +0.14; CRP/Fe, -0.08; CRP/NeuTOX, +0.23; MonATT, +0.22. On the contrary, the MR analysis did not show any relationship between CRP and other markers (R2: 0.05 for CRP ≥ 0.30 mg/dL;; 0.02 for CRP ≤ 0.29 mg/dL). ROC analysis of the parameters yielded the following results: NeuSeg is a moderately accurate inflammation marker with Area Under the Curve (AUC) of 0.71. The other parameters are less accurate markers of inflammation (AUC) compared to CRP: WBC, 0.70; Fib, 0.67; Alb, 0.64; Fe, 0.64; Band, 0.59. The markers with the best combination of Sensitivity (SS) and Specificity (SP) were: Fib (SS, 52.7; SP, 77.5 for 400 mg/dL cut-off) and Band (SS, 17.6; SP, 98.0 for 0.3 K/μL cut-off). The correlation between CRP and all the parameters studied, except Linf REA, is significant but low because they are affected by many conditions aside from inflammation. None of them is able to predict CRP values. The diagnostic accuracy of each single inflammatory marker is lower in comparison to CRP. In order to increase the diagnostic accuracy of inflammation markers, an evaluation of several parameters simultaneously is warranted, particularly in the absence of CRP measurement

Evaluation of Erythrocytes, Platelets, and Serum Iron Profile in Dogs with Chronic Enteropathy

The aim of this study is to evaluate iron status, erythrocyte, and platelet modifications in dogs with chronic enteropathy (CE). Dogs were grouped as food-responsive diarrhea (FRD, n = 11), antibiotic-responsive diarrhea (ARD, n = 5), and steroid-responsive diarrhea (SRD, n = 6) relating to therapeutic-response. Clinical and haematological findings, evidence of gastrointestinal blood loss, and iron metabolism were evaluated before and after treatment. A mild normocytic or microcytic anemia and thrombocytosis were identified, respectively in 18.0% and 31.8% of CE dogs. No significant differences between pre- and posttreatment of hematocrit, haemoglobin, and mean corpuscular volume, platelet count and mean platelet volume were found. Statistical analysis pointed out significant differences between pre- and posttreatment in serum iron (P < .03) and unsaturated iron binding capacity (UIBC) (P < .01). No significant correlations were found between these parameters and canine Inflammatory Bowel Disease activity index and pattern of CE as well

Epidemiology of Breed-Related Mast Cell Tumour Occurrence and Prognostic Significance of Clinical Features in a Defined Population of Dogs in West-Central Italy

Abstract: Canine mast cell tumours (MCTs) present a wide variety of challenging clinical behaviours in terms of predicting the prognosis and choosing appropriate treatment. This study investigated the frequency, risk, and prognostic factors of MCTs in dogs admitted to a single veterinary teaching hospital (VTH). Breed, age, sex, and sexual status in ninety-eight dogs with MCTs (MCT-group) were compared with a control group of 13,077 dogs (VTH-group) obtained from the VTH clinical database from January 2010 to January 2016. Within the MCT-group, signalment, location, size, mass number, ulceration, histopathological grading, presence of lymph node, or distant metastases were compared with each other and with the outcome. Boxers (OR 7.2), American Pit Bull Terriers (OR 5.4), French Bulldogs (OR 4.4) and Labrador Retrievers (OR 2.6) were overrepresented. The MCT-group was significantly older than the VTH-group (p &lt; 0.0001). In comparison with the VTH group, in the MCT-group neutered dogs (OR 2.1) and spayed females (OR 2.3) were predominant compared to intact dogs and intact females, respectively. Ulceration (OR 5.2) and lymph node metastasis (OR 7.1) occurred more frequently in larger MCTs. Both ulceration and MCTs &gt; 3 cm were highly associated with lymph node metastasis (OR 24.8). Recurrence was associated with MCT-related death (OR 10.50, p = 0.0040), and the latter was associated with shorter survival times (p = 0.0115). Dogs with MCTs &gt; 3 cm (p = 0.0040), lymph node metastasis (p = 0.0234), or elevated WHO stage (p = 0.0158) had shorter survival times. A significantly higher frequency of MCTs was found in specific breeds, and in older and neutered dogs. MCTs &gt; 3 cm and lymph node or distant metastases were associated with shorter survival times