Evaluation of the effectiveness of Duddingtonia flagrans and Monacrosporium thaumasium in the biological control of gastrointestinal nematodes in female bovines bred in the semiarid region

Brazil has a herd of 212 million cattle and 171 million hectares of pastures that produce approximately 96 % of Brazilian beef. The Brazilian production system enables animal infection by endoparasites, which are considered one of the main obstacles for the development of this industry and are responsible for considerable economic losses. The control of parasitic diseases is performed via the administration of antiparasitic drugs, but they leave residues of the products in the treated animal, affect non-target organisms and select resistant strains of the parasites. The species D. flagrans and M. thaumasium are promising and sustainable alternatives for controlling gastrointestinal helminths of ruminants and other herbivores. In this study, we evaluated the efficacy of isolates of these species, formulated in a sodium alginate matrix and administered twice a week, to reduce the number of environmental infective larvae of gastrointestinal nematodes that affect prepubescent zebu females. The treated animals presented fewer eggs and a lower number of infective larvae per gram of faeces (p?<?0.05). The pastures occupied by treated animals showed a statistically significant reduction (p?<?0.05) of the number of L₃ and, furthermore, the genera Cooperia sp., Haemonchus sp., and Oesophagostomum sp. were the most prevalent. The average weight of the animals did not differ statistically (p?>?0.05) among the treated and control groups. The use of sodium alginate pellets as vehicle for delivery of the fungus mycelia D. flagrans (isolate AC001) and M. thaumasium (isolate NF34A) proved effective in controlling trichostrongylids in prepubescent cows bred in the semi-arid region, with an effective reduction in the number of infective larvae in the pastures.     

Prevalence and risk factors associated with goat gastrointestinal helminthiasis in the Sertão region of Paraíba State,Brazil

Gastrointestinal helminthiasis represents an obstacle to goat raising, causing severe damage to herds such as growth retardation, weight loss, and even death. In this study, we aimed to determine the prevalence and risk factors associated to goat gastrointestinal helminthiasis in the Sertão region of Paraíba State, Brazil. A total of 256 goats from 54 farms were systematically sampled. Blood and fecal samples were collected from each animal for egg per gram (EPG), larval culture, and packed cell volume (PCV) analyses. We found that 79.3 % of the goats investigated were parasitized with gastrointestinal helminths. Significant correlation (p?=?0.004) was observed between the EPG and PCV of the animals studied, and it was observed that the EPG increases as the PCV decreases. In the larval culture, the most prevalent helminth was Haemonchus sp. (83.2 %). Age and sex were significant variables (p?≤?0.20) for the development of gastrointestinal helminths: 86.8 % of animals over 36 months of age and 81.7 % of females were infected. The variable type of animal exploitation was also significant, with 90.3 % (p?≤?0.20) of the animals presenting double suitability (milk and meat). The Sertão region of Paraíba State presents high prevalence of gastrointestinal helminthiasis in goats, and age and type of animal exploitation are the most relevant risk factors to the development of these parasites.     

Prevalence of antimicrobial resistance in enteric Escherichia coli from domestic pets and assessment of associated risk markers using a generalized linear mixed model

Antimicrobial resistance (AMR) is a growing global public health problem, which is caused by the use of antimicrobials in both human and animal medical practice. The objectives of the present cross-sectional study were as follows: (1) to determine the prevalence of resistance in Escherichia coli isolated from the feces of pets from the Porto region of Portugal against 19 antimicrobial agents and (2) to assess the individual, clinical and environmental characteristics associated with each pet as risk markers for the AMR of the E. coli isolates.     

Atypical fibrosarcoma in the skin of a Roborovski hamster (Phodopus roborovskii)

A 2.5‐year‐old intact male Roborovski hamster (Phodopus roborovskii) was presented with a large subcutaneous mass overlying the abdomen, affecting the animal's ambulation and access to different compartments of the cage through narrow tubing. Ultrasound examination delineated a well‐circumscribed mass in the subcutis of the caudoventral abdominal region. The mass was surgically excised and on cytologic examination showed, in a background of blood, a small population of individually arranged oval to spindle‐shaped cells that exhibited a moderate degree of anisokaryosis, coarsely stippled chromatin, one or more prominent nucleoli, and lightly basophilic well‐defined cytoplasmic processes. Histologically, the mass was composed of interlacing streams and bundles of pleomorphic spindle cells (ganglion‐like cells) with variable amounts of collagenous stroma. The neoplastic cells exhibited moderate features of malignancy. These cells stained intensely with vimentin, but not with any other markers, including antibodies to cytokeratin AE1/AE3, S100 protein, desmin, smooth muscle actin, synaptophysin, neurofilament, and androgen receptor. Based on histologic features, the mass was diagnosed as an atypical fibrosarcoma. This is the first report of an atypical fibrosarcoma in a Roborovski hamster and one of few reports of atypical fibrosarcoma in domesticated hamsters overall.     

Genomic analysis of influenza A virus from captive wild boars in Brazil reveals a human-like H1N2 influenza virus

Influenza is a viral disease that affects human and several animal species. In Brazil, H1N1, H3N2 and 2009 pandemic H1N1 A(H1N1)pdm09 influenza A viruses (IAV) circulate in domestic swine herds. Wild boars are also susceptible to IAV infection but in Brazil until this moment there are no reports of IAV infection in wild boars or in captive wild boars populations. Herein the occurrence of IAV in captive wild boars with the presence of lung consolidation lesions during slaughter was investigated. Lung samples were screened by RT-PCR for IAV detection. IAV positive samples were further analyzed by quantitative real-time PCR (qRRT-PCR), virus isolation, genomic sequencing, histopathology and immunohistochemistry (IHC). Eleven out of 60 lungs (18.3%) were positive for IAV by RT-PCR and seven out of the eleven were also positive for A(H1N1)pdm09 by qRRT-PCR. Chronic diffuse bronchopneumonia was observed in all samples and IHC analysis was negative for influenza A antigen. Full genes segments of H1N2 IAV were sequenced using Illumina's genome analyzer platform (MiSeq). The genomic analysis revealed that the HA and NA genes clustered with IAVs of the human lineage and the six internal genes were derived from the H1N1pdm09 IAV. This is the first report of a reassortant human-like H1N2 influenza virus infection in captive wild boars in Brazil and indicates the need to monitor IAV evolution in Suidae populations.     

Evidence for widespread Leishmania infantum infection among wild carnivores in L. infantum periendemic northern Spain

Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n = 53), 29% of foxes (n = 48), 29% of stone martens (n = 21) and in 25–50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Acupuncture Diagnosis in Equine Stifle Disease

The stifle is a large and complex joint in quadrupeds, including horses, making the diagnosis of stifle diseases a challenge for the clinician. The detection of “stifle syndrome” during acupuncture examination of the equine stifle may be useful for diagnosis of stifle diseases in horses. The aim of the present study was to describe stifle syndrome diagnosis in horses. From 2001 through 2012, 810 athletic horses involved in different disciplines were examined using the technique of acupuncture diagnosis. Stifle syndrome was suspected when the following points were reactive: the sacral point suggestive of stifle disease located at the sacral origin of the biceps femoris muscle, bladder (BL)-20, BL-21, and stomach (ST)-30, together with the three local points in the muscle groove of the posterolateral aspect of the thigh. The diagnostic protocol included radiography and/or ultrasonography in certain cases. The horses were evaluated because of reduced athletic performance, routine or prepurchase examination, hind limb lameness, refusal to jump, inappetence, upward fixation of the patella, thoracolumbar spine or sacroiliac pain, suspensory lesion, and front hoof problems, and 86 (9.4%) were diagnosed with stifle syndrome. The sacral acupuncture point suggestive of stifle syndrome correlated with BL-21, local stifle points, gall bladder (GB)-27, spleen-13, BL-23, BL-52, BL-26, and/or GB-25, ST-30, BL-54 (P < .0001), and BL-20 (P = .003). Twenty-two (25.6%) of the 86 horses with stifle syndrome underwent ultrasonography and/or radiography, and lesions were detected in 21. In conclusion, acupuncture for diagnosis of stifle disease may be considered a diagnostic method to be included in routine clinical examination of horses, and its reproducibility shall be investigated in further studies.     

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Ultrastructural morphological injuries and maturation rates were investigated in equine oocytes exposed to vitrification solutions (VS) containing synthetic ice blockers (SIBs) during different exposure times. In experiment 1, compact cumulus-oocyte complexes (COCs; n = 30) were randomly allocated to treatments: (1) fresh fixed (control); (2) VS-1 (1.4 M dimethyl sulfoxide [DMSO] + 1.8 M ethylene glycol [EG] + 1% SIB) for 3 minutes of equilibrium time and VS-2 (2.8 M DMSO + 3.6 M EG + 0.6 M sucrose + 1% SIB) for 1 minute (Eq-long); and (3) VS-1 for 1.5 minutes and VS-2 for 30 seconds (Eq-short). In experiment 2, compact (n = 248) and expanded (n = 264) COCs were evenly distributed to the following treatments: (1) immediate maturation in vitro (control); (2) vitrification using the Eq-short protocol as in experiment 1; and (3) vitrification using a stock solution containing 2.8 M formamide, 2.8 M DMSO, 2.7 M EG, 7% polyvinylpyrrolidone, and 1% SIB (Eq-short-mod). More (P < .02) oocytes with normal ultrastructural morphology were seen in fresh control and Eq-short groups than in Eq-long group. Metaphase-II (MII) rates were higher (P < .05) for oocytes with expanded cumulus than compact cumulus in the control group, and higher (P < .05) for oocytes with expanded cumulus than compact cumulus in Eq-short and Eq-short-mod groups. No difference in MII rates was detected among groups within each type of COC. In conclusion, reduction of exposure time to VS better preserved oocyte ultrastructural features, and MII rates were higher for vitrified oocytes with expanded cumulus. This study advances our knowledge on potential alternatives for vitrification of immature equine oocytes.     

Comparison of Cryoprotective Effects of Lycopene and Cysteamine in Different Cryoprotectants on Bull Semen and Fertility Results

The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty‐four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5 mm or lycopene 500 μg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non‐return rates of inseminated cows. However, this result was not considered to be statistically important.