Stem cell factor supports migration in canine mesenchymal stem cells

Adult Mesenchymal Stem Cells (MSC) are cells that can be defined as multipotent cells able to differentiate into diverse lineages, under appropriate conditions. These cells have been widely used in regenerative medicine, both in preclinical and clinical settings. Initially discovered in bone marrow, MSC can now be isolated from a wide spectrum of adult and foetal tissues. Studies to evaluate the therapeutic potential of these cells are based on their ability to arrive to damaged tissues. In this paper we have done a comparative study analyzing proliferation, surface markers and OCT4, SOX9, RUNX2, PPARG genes expression in MSC cells from Bone marrow (BMMSC) and Adipose tissue (ASC). We also analyzed the role of Stem Cell Factor (SCF) on MSC proliferation and on ASCs metalloproteinases MMP-2, MMP-9 secretion. Healthy dogs were used as BMMSC donors, and ASC were collected from omentum during elective ovariohysterectomy surgery. Both cell types were cultured in IMDM medium with or without SCF, 10% Dog Serum (DS), and incubated at 38 °C with 5% CO2. Growth of BMMSCs and ASCs was exponential until 25–30 days. Flow citometry of MSCs revealed positive results for CD90 and negative for CD34, CD45 and MCH-II. Genes were evaluated by RT-PCR and metalloproteinases by zymografy. Our findings indicate morphological and immunological similarities as well as expression of genes from both origins on analyzed cells. Furthermore, SCF did not affect proliferation of MSCs, however it up-regulated MMP-2 and MMP-9 secretion in ASCs. These results suggest that metalloproteinases are possibly essential molecules pivoting migration.     

Risk factors associated with feline urolithiasis

Urinary tract diseases are among the main reasons for consultation in veterinary clinics and hospitals. It affects animals of any age, breed and gender. Among the diseases that affect this system, urolithiasis is the second largest cause of clinical signs compatible with feline urinary tract disease. The term urolithiasis refers to the presence of uroliths in any region of the urinary tract, but it is more commonly seen in the bladder and urethra. Uroliths are classified based on the type of mineral present in their composition, therefore, quantitative and qualitative analyzes are important for a better therapeutic approach. The animals may suffer from the disease and be asymptomatic, or show nonspecific clinical signs, making the diagnosis difficult. The disease should not be seen as a single problem, but as a consequence of various disorders. As dietary, metabolic, genetic and infectious causes, as well as factors that potentiate the chance of development of uroliths such as breed, age, sex, age range, obesity, sedentary lifestyle, geographic region and climate. Thus, the knowledge of the factors that influence the formation of uroliths, as well as the understanding of the pathophysiology, are key elements for better alternatives of therapy and prevention. The recognition of these factors helps to identify susceptible populations, minimizing exposure and increasing the protection factors, which facilitates the diagnosis and treatment of patients with urolithiasis. The objective of this paper is to present the main risk factors involved in the formation of urinary lithiasis in felines.     

Genomic characterization of the first oral avian papillomavirus in a colony of breeding canaries (<Emphasis Type="Italic">Serinus canaria</Emphasis>)

Papillomaviruses are non-enveloped, DNA viruses that infect skin and mucosa of a wide variety of vertebrates, causing neoplasias or simply persisting asymptomatically. Avian papillomaviruses, with six fully sequenced genomes, are the second most studied group after mammalian papillomaviruses. In this study, we describe the first oral avian papillomavirus, detected in the tongue of a dead Yorkshire canary (Serinus canaria) and in oral swabs of the same bird and other two live canaries from an aviary in Madrid, Spain. Its genome is 8,071 bp and presents the canonical papillomavirus architecture with six early (E6, E7, E1, E9, E2, E4) and two late open reading frames (L1 and L2) and a long control region between L1 and E6. This new avian papillomavirus L1 gene shares a 64% pairwise identity with FcPV1 L1, so it has been classified as a new species (ScPV1) within the Ethapapillomavirus genus. Although the canary died after showing breathing problems, there is no evidence that the papillomavirus caused those symptoms so it could be part of the oral microbiota of the birds. Hence, future investigations are needed to evaluate the clinical relevance of the virus.     

B-mode ultrasound examination of canine mammary gland neoplastic lesions of small size (diameter &lt; 2 cm)

Ultrasonography is a valuable tool for the evaluation of neoplastic lesions in the dog and there is a growing interest in the use of this technique for the stadiation of canine mammary tumours. An accurate assessment of small sized nodules facilitates the stadiation of the mammary lesions and helps the clinician in the choice of the most indicated surgical therapy. The aim of this study was to identify those ultrasound criteria that may be useful in discriminating between benign and malignant lesions of small size (diameter smaller than 2 cm). Sixty-two nodules, < 2 cm in larger diameter, belonging to thirty-five bitches presented between January 2012 and February 2014 were evaluated. Tumours were observed by conventional ultrasound and assessed for: shape (regular-irregular), limit (defined-ill-defined), margins (regular-irregular), echogenicity (hypoechoic-isoechoic-hyperecoic), echotexture (homogeneus-heterogeneus), presence of hyperecoic halo, distal acoustic enhancement or shadowing and surrounding tissue alterations. Among the alterations in surrounding tissues, the disruption of the glandular tissue and the increase in echogenicity of the peritumoral tissues were assessed. Thereafter, bitches were subjected to mastectomy and nodules were evaluated histologically. None of the ultasound criteria considered in the current study showed a statistically significant relation with malignancy, except for the presence of alterations in the tissue surrounding the nodules. According to our results, this characteristic may indicate malignancy, however its subjectivity may affect the applicability in clinical practice. In conclusions, conventional ultrasound in bitches had a limited ability in discriminating benign and malignant mammary gland neoplastic lesions of small size (diameter?<?2 cm).     

Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.     

The IMPDH inhibitors,ribavirin and mycophenolic acid,inhibit peste des petits ruminants virus infection

Peste des petits ruminants virus (PPRV) causes highly contagious diseases in domestic and particular wild small ruminants, leading to substantial economic loss. The development of effective and cheap antiviral medications shall help to circumvent this emerging burden. In this study, we found that ribavirin, a competitive inosine-5’-monophosphate dehydrogenase (IMPDH) inhibitor, significantly inhibits the replication of PPRV. As IMPDH is a key enzyme in purine nucleotide synthesis, supplementation of exogenous guanosine attenuate the anti-PPRV effect of ribavirin. Interestingly, an uncompetitive IMPDH inhibitor, mycophenolic acid (MPA), exerted more potent antiviral effect again PPRV. Similarly, this effect was largely restored upon supplementation of guanosine. Thus, we have demonstrated that the IMPDH inhibitors ribavirin and MPA combat PPRV infection through purine nucleotide depletion. Because both regimens have been widely used in the clinic for treating viral infection or organ rejection in transplantation patients for decades, respectively, repurposing these existing safe and cheap medications may provide a new avenue for combating PPRV infection.     

Knowledge,attitudes and practices towards <Emphasis Type="Italic">cystic echinococcosis</Emphasis> in livestock among selected pastoral and agro-pastoral communities in Uganda

A cross-sectional study was done from March 2013 to May 2014 to assess knowledge, attitudes, and practices towards cystic echinococcosis (CE) or hydatidosis among selected pastoral and agro-pastoral communities in Uganda. A structured questionnaire was administered to 381 respondents. Multivariate logistic regression analysis was done to find the relationship between knowledge about CE and factors such as age, sex, and level of education across all regions. The odds ratio and confidence interval were used to determine the difference in responses across regions. It was shown that age above 36 years was significantly (p < 0.001) associated with awareness about CE in livestock. Likewise, uneducated (p < 0.0001) and agro-pastoralists (p = 0.01) were significantly less knowledgeable than the educated and pastoralists across all regions. The overall knowledge towards CE in livestock was low 17.8% (95% CI = 14.0–21.6). Dog ownership was high and they never dewormed their freely roaming dogs. Dogs shared water with livestock. In conclusion, knowledge about CE in livestock was low across all regions. Therefore, public health education and formulation of policies towards its control by the relevant stakeholders should be done. Also, the true prevalence of CE in livestock needs to be done so that the magnitude and its public health significance are elucidated.     

Drivers,challenges and opportunities of forage technology adoption by smallholder cattle households in Cambodia

Forage technology has been successfully introduced into smallholder cattle systems in Cambodia as an alternative feed source to the traditional rice straw and native pastures, improving animal nutrition and reducing labour requirements of feeding cattle. Previous research has highlighted the positive impacts of forage technology including improved growth rates of cattle and household time savings. However, further research is required to understand the drivers, challenges and opportunities of forage technology for smallholder cattle households in Cambodia to facilitate widespread adoption and identify areas for further improvement. A survey of forage-growing households (n = 40) in July–September 2016 examined forage technology adoption experiences, including reasons for forage establishment, use of inputs and labour requirements of forage plot maintenance and use of forages (feeding, fattening, sale of grass or seedlings and silage). Time savings was reported as the main driver of forage adoption with household members spending approximately 1 h per day maintaining forages and feeding it to cattle. Water availability was reported as the main challenge to this activity. A small number of households also reported lack of labour, lack of fencing, competition from natural grasses, cost of irrigation and lack of experience as challenges to forage growing. Cattle fattening and sale of cut forage grass and seedlings was not found to be a widespread activity by interviewed households, with 25 and 10% of households reporting use of forages for these activities, respectively. Currently, opportunities exist for these households to better utilise forages through expansion of forage plots and cattle activities, although assistance is required to support these households in addressing current constraints, particularly availability of water, if the sustainability of this feed technology for smallholder cattle household is to be established in Cambodia.     

Brucellosis in migratory sheep flock from Maharashtra,India

Brucellosis is a zoonotic disease worldwide distributed and having the economic as well as public health importance. The prevalence of brucellosis among sheep flock having history of abortions was studied. A total of 229 samples comprising of 157 blood and 72 clinical samples (vaginal swabs) were collected from 157 animals. Clinical samples were processed for the isolation of Brucella melitensis. Serum samples (n = 157) were tested by Rose Bengal plate test (RBPT) and i-ELISA. A total of 68 (43.31%) and 104 (66.24%) samples were positive by RBPT and ELISA, respectively. Brucella isolates (n = 2) were recovered from clinical samples. Both isolates demonstrated amplification for bcsp 31 and IS711 genes. On AMOS PCR, both the isolates amplified at 731 bp, i.e., belongs to B. melitensis species. The incidence of B. melitensis in a migratory flock warns the thorough testing and culling of Brucella-infected sheep from the flock on a continuous basis; otherwise, such incidence will be routine and poor farmers will be at a loss.