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The present study describes the morphological and immunohistochemical characteristics of a case of diffuse seminoma in a 16-year-old male mixed-breed horse. According to the owner, the animal's left testicle had been gradually increasing in size over a period of 2 months. On palpation, the testicle had a firm consistency, with no sensitivity to digital pressure, was adhered to the scrotum and measuring 16 cm × 8 cm. In the ultrasound examination, it presented a heterogeneous texture and areas of hypoechogenic echogenicity without visualization of the mediastinum. Therefore, the bilateral orchiectomy was performed. After the surgical procedure, it was found that the affected testicle presented a firm mass measuring 9 cm × 7 cm × 3.5 cm. Histologically, a multilobulated, non-encapsulated and invasive tumour mass was found, which replaced the seminiferous tubules, consisting of polygonal cells arranged in a mantle that varied from cohesive to loosely cohesive, supported by a scarce fibrous stroma. In the immunohistochemical examination, the neoplastic cells showed positive immunolabelling for OCT4 and C-KIT. In this report, the physical examination combined with the ultrasonographic examination were fundamental to the therapeutic management of the case, and the final diagnosis was made after histopathological and immunohistochemical tests., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

An analytical procedure for the multielement determination in enteral nutrition formulations employing slurry sampling and inductively coupled plasma optical emission spectrometry (ICP OES) is proposed. A two-level full-factorial design was applied to assess the influence of the presence of stabilizing agents (HNO 3 , Triton X-100 and ethanol) on the composition of the slurry. Multiple response was established as a dependent variable. The experimental conditions for the preparation of the slurry were: 2.0 mL of sample and 8.0 mL of 10% (v/v) HNO 3 . The limits of detection (LOD) were 5; 9; and 10 µg L -1 for Cu, Fe, Zn, respectively. For P, and K, the LOD were 8 and 24 mg L -1 , respectively. The method was applied for the analysis of three enteral nutrition formulation samples and the obtained concentrations ranges were (in mg L -1 ): 0.41-0.43 (Cu), 2.0-2.9 (Fe), 1.7-3.1 (Zn), 682-1409 (K), and 217-344 (P)., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Loss of genetic integrity can occur during the long-term conservation of seeds. We have studied these effects in seeds of rice (Oryza sativa L.) and common bean (Phaseolus vulgaris L.) exposed to accelerated aging (elevated temperature and moisture) conditions. Tests of first count, germination, and germination speed index were performed to measure physiological quality; cytogenetic tests and comet assay were used to evaluate genetic integrity. With aging, we observed a decrease in mitotic index and an increase in the frequency of chromosomal alterations in root cells of imbibed seeds, as well as increased DNA damage (comet assay) in dry and imbibed seed embryos of both species. The comet assay can be a useful technique for measuring genetic integrity in seed conservation programs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Leptospirosis is a disease that negatively affects the productive and reproductive indices of ruminants. Sheep are considered highly resistant to infection, although susceptibility may vary among breeds. Thus, the aim of the present study was to analyze the susceptibility between sheep breeds to the experimental infection by leptospires of the Pomona serogroup. Pomona serogroup, Kennewicki serovar strain (1 × 107 bacteria) was inoculated via the conjunctival route in 12 sheep divided into two groups, one comprising Santa Inês ewes and the other comprising crossbred sheep. In each group, five ewes were challenged with the bacterial strain and one was used as control. All sheep were monitored for 60 days, during which blood samples were collected for serological diagnosis and urine and vaginal fluid samples for molecular and microbiological analyses. Finally, as ewes were submitted to euthanasia and necropsy, some tissues of interest were collected for microbiological, molecular, and histopathological diagnoses. The groups were compared regarding the number of positive reactions according to diagnostic tests. All sheep in each group presented antibodies to Leptospira in all serological analyses, except animals of the control group. However the Santa Inês sheep presented higher concentration and duration of the titers, and their positive reactions were detected earlier than those in crossbred sheep. The antibody titers in group A (median 200, geometric mean 317.48) were significantly different from the group B (median 800, geometric mean 918.96) at D60 post-infection (P = 0,032). The Santa Inês sheep presented a higher number of positive reactions than did the crossbred sheep in the molecular diagnostic tests. According to the molecular diagnosis, the Santa Inês sheep presented more reactions (urine and vaginal fluid) compared to crossbred ewes, but there was no predominance in the detection of leptospiral DNA when comparing urine and vaginal fluid results, nor even between the number of positive kidneys and uterus. The Santa Inês sheep presented a higher number of positive bacteriological cultures. No sheep in either group presented alterations in anatomopathological and histopathological findings. Pure-bred sheep may be more susceptible than crossbred sheep to infection by Leptospira sp. Our findings emphasize the importance of the genital tract as a site of extraurinary infection and indicate the possibility of venereal transmission in the species., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Three spectrophotometric methods were developed for the determination of copper (Cu) in sugar cane spirit using the chromogenic reagents neocuproine, cuprizone, and bathocuproine. Experimental conditions, such as reagent concentration, reducer concentration, pH, buffer concentration, the order of addition of reagents, and the stability of the complexes, were optimized. The work range was established from 1.0 to 10.0 µg/mL, with correlation coefficients of >0.999 for all three optimized methods. The methods were evaluated regarding accuracy by addition and recovery tests at five concentration levels, and the obtained recoveries ranged from 91 to 105% (n = 3). Precision was expressed as RSD (relative standard deviation), with values ranging from 0.01 to 0.17% (n = 10). The method using the chromogenic reagent cuprizone presented the greatest molar absorptivity, followed by bathocuproine and neocuproine. The methods were applied for the determination of Cu in sugar cane spirit, and the results were compared with a reference method by flame atomic absorption spectrometry (FAAS). Calibration curve solutions for FAAS analysis were prepared in a 40% (v/v) alcohol medium in a range of concentrations from 0.5 up to 5 µg/mL. Measurements for Cu determination were carried out at a wavelength of 324.7 nm. The concentrations obtained for Cu in sugar cane spirit samples from Brazil were between 1.99 and 12.63 µg/mL, and about 75% of the samples presented Cu concentrations above the limit established by Brazilian legislation (5.0 µg/mL or 5.0 mg/L).

This study aimed to assess the feasibility of comet and cytogenetic tests as tools for evaluating genomic instability in seeds of Oryza sativa L. (rice) and Phaseolus vulgaris (beans) L. from gene banks. Rice and beans were exposed to methyl methanesulfonate (MMS) as a reference DNA damaging agent. Seeds of two accessions of rice and beans were obtained from Embrapa Rice and Beans - Brazil. Seed groups were imbibed in three concentrations of MMS for three periods of time to carry out cytogenetic tests, and for one period for the comet test. At concentrations of 10 and 15 mg/L, MMS induced cytotoxic and/or mutagenic effects in the meristematic cells of roots from all the accessions of both species. In the comet test, MMS induced genotoxic effects at all the concentrations in the evaluated accessions of rice and beans, except in one accession of beans at the lowest concentration (5 mg/L). Both species showed sensitivity to MMS. The comet test can be proposed for the measurement of genomic instability in accessions of rice and beans in gene banks, as being more sensitive than the cytogenetic tests used.

This study reports the clinicopathological, immunohistochemical, and molecular findings from two cases of systemic toxoplasmosis in pigs showing apathy and dyspnea. In the post-mortem examination, severe diffuse necrotizing bronchointerstitial pneumonia with numerous intralesional tachyzoites of Toxoplasma gondii was observed. The lungs had not collapsed but were diffusely reddened, and the parenchyma showed friable whitish subpleural nodules with multifocal to coalescent distribution and diameters of 0.5-1.0 cm. The histopathological findings comprised mononuclear inflammation and multifocal areas of necrosis in alveolar septa (cases 1 and 2). In addition, esophagitis and ulcerations in the mucosa of the stomach and the small and large intestines were observed (case 1). Immunohistochemical analysis using anti-T. gondii antibodies on lung tissue in both cases revealed strong immunolabeling of free tachyzoites and tachyzoites in the cytoplasm of histiocytes and in cysts. Nested PCR targeting a 155-bp fragment of the B1 gene of T. gondii was positive for the DNA extracted from lung fragments from the two pigs. Genotyping of the samples by means of PCR-RFLP (10 markers) and by means of microsatellites (15 of them) revealed that these animals were infected with T. gondii that was molecularly characterized as the non-archetypal genotype Chinese 1. This presents worldwide circulation, but it had not previously been described in Brazil. The microsatellite analysis showed that the animals were infected with the same T. gondii isolate circulating in the environment.

Our study describes the clinical, epidemiologic, pathologic, immunohistochemical, and molecular aspects PCR of a case of cutaneous pythiosis in a donkey (Equus asinus) from Brazil. During a dry period, the animal grazed for 4 months around a pond where the vegetation remained green. Skin lesions were nodular, multifocal, and disseminated, mainly involving the legs, ventral chest, and mammary gland. On cut surface, there were multifocal to coalescent discrete yellow foci, and occasional small cavitations with a few kunkers. Ulcerative nodular pyogranulomatous and eosinophilic dermatitis with folliculitis and furunculosis were observed histologically. Hyphae were observed in sections stained with Gomori methenamine silver. Immunohistochemistry with Pythium insidiosum antibodies yielded strong immunostaining of hyphae. P. insidiosum DNA was extracted from tissues in paraffin blocks by amplification of a fragment of 105 bp, which targets the 5.8S ribosomal gene. After the diagnosis of pythiosis, the larger skin lesions were excised and treated as second intention healing wounds, which were completely healed 30 days after resection. Small skin lesions regressed spontaneously in ~60 days. The granulomatous inflammation and outcome of the disease in this donkey were similar to cases of pythiosis in cattle., (© 2016 The Author(s).)

An assisted liquid-liquid extraction of copper, iron, nickel and zinc from vegetable oil samples with subsequent determination by high-resolution continuum source flame atomic absorption spectrometry (HR-CS FAAS) was optimized by applying a full factorial design in two levels and the response surface methodology, Box-Behnken. The effects of the acid concentration and the amplitude, cycle and time of sonication on the extraction of the analytes, as well as their interactions, were assessed. In the selected condition (sonication amplitude = 66%, sonication time = 79 s, sonication cycle = 74%), using 0.5 mol L(-1) HCl as the extractant, the limits of quantification were 0.14, 0.20, 0.21 and 0.04 μg g(-1) for Cu, Fe, Ni and Zn, respectively, with R.S.D. ranging from 1.4% to 3.6%. The proposed method was applied for the determination of the analytes in soybean, canola and sunflower oils., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The cashew nut releases a substance that is known as cashew nut shell liquid (CNSL). There are both natural (iCNSL) and technical (tCNSL) cashew nut shell liquids. This study used an Artemia salina bioassay to evaluate the toxic effects of iCNSL and tCNSL cashew nut shell liquids. It also evaluated the toxicity, cytotoxicity, and mutagenicity of CNSL and its effects on the damage induced by copper sulfate (CuSO4·5H2O) on the meristems' root of Allium cepa. Effects of the damage induced by CuSO4·5H2O were evaluated before (pre-), during (co-), and after (post-) treatments. The iCNSL contained 94.5% anacardic acid, and the tCNSL contained 91.3% cardanol. The liquids were toxic to A. salina. Toxicity, cytotoxicity, and mutagenicity were observed with iCNSL compared with the negative control. Similarly, iCNSL failed to inhibit the toxicity and cytotoxicity of CuSO4·5H2O. The tCNSL was not toxic, cytotoxic, or mutagenic in any of the concentrations. However, the lowest iCNSL concentrations and all of the tCNSL concentrations had preventive, antimutagenic, and reparative effects on micronuclei and on chromosomal aberrations in the A. cepa. Therefore, protective, modulating, and reparative effects may be observed in the A. cepa, depending on the concentration and type of CNSL used.

Two experiments, each with 10 pregnant ewes (8 treated and 2 controls) were performed to determine if nursing lambs of lactating ewes become intoxicated when the ewes ingest Ipomoea asarifolia but do not show clinical signs themselves. In the first experiment the sheep grazed I. asarifolia in the field while in the second, sheep were maintained in individual bays consuming dry I. asarifolia at 10% and 20% into their ration. In both experiments the lambs remained confined, consuming only their mother's milk. Four of 8 lambs in the grazing experiment and the 4 nursing lambs from the ewes given 20% I. asarifolia showed signs of I. asarifolia poisoning. These results confirm that the tremorgenic compound of I. asarifolia or its toxic metabolites are eliminated in milk and can intoxicate nursing lambs., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The current article describes outbreaks of vesicular stomatitis (VS) in horses and cattle in Paraiba and Rio Grande do Norte states, northeastern Brazil, between June and August 2013. The reported cases affected 15-20 horses and 6 cattle distributed over 6 small farms in 4 municipalities, but additional data indicated the involvement of a large number of animals on several farms. The disease was characterized by blisters; eruptive lesions in coronary bands, lips, mouth, and muzzle; salivation; claudication and loss of condition. Swollen lower limbs and lips, and ulcerated and erosive areas in the lips and muzzle were observed in some horses. A necrotizing vesiculopustular dermatitis and stomatitis was observed histologically. Vesicular stomatitis virus was isolated from the vesicular fluid of a horse lesion and shown to be serologically related to the VS Indiana serogroup (VSIV) by virus neutralization. Convalescent sera of affected horses and cattle, and from healthy contacts, harbored high levels of neutralizing antibodies against the isolated virus (named VSIV-3 2013SaoBento/ParaibaE). Genomic sequences of VSIV subtype 3 (Vesicular stomatitis Alagoas virus) were amplified by reverse transcription polymerase chain reaction out of clinical specimens from a cow and a horse from different farms. Nucleotide sequencing and phylogenetic analysis of the phosphoprotein gene indicated that the 2 isolates were derived from the same virus and clustered them in VSIV-3, along with VS viruses identified in southeastern and northeastern Brazil in the last decades. Thus, the present report demonstrates the circulation of VSIV-3 in northeastern Brazil and urges for more effective diagnosis and surveillance., (© 2014 The Author(s).)

We describe the pathologic and immunohistochemical findings in five cases of rabies in the insectivorous Pallas's mastiff bat (Molossus molossus) in the city of Patos, state of Paraíba, northeastern Brazil. All cases were found during the day, in different neighborhoods, prostrate and unable to fly. No significant lesions were observed at necropsy or on histologic examination. The brains of the five bats showed strong immunohistochemical labeling for rabies virus in the form of large corpuscles or multiple clusters of granules within the perikarya of neurons in different areas of the brain, mainly in the cerebral cortex and cerebellar Purkinje cells. Three bats tested by direct immunofluorescence and mouse inoculation were also positive for rabies. These data demonstrate that M. molossus may be infected with the rabies virus and develop rabies, even without histologic lesions in the central nervous system, and can be a source of infection for humans and domestic animals.

Nasal and cutaneous aspergillosis is reported in an adult goat. The clinical signs were severe respiratory distress due to partial nasal obstruction, bilateral mucopurulent nasal discharge, skin nodules on the ears and dorsal nasal region and focal depigmentation of the ventral commissure of the right nostril. At necropsy examination, sagittal sectioning of the head revealed a yellow irregular mass extending from the nasal vestibule to the frontal portion of the nasal cavity. Microscopically, there was pyogranulomatous rhinitis and dermatitis, with numerous intralesional periodic acid-Schiff-positive fungal hyphae morphologically suggestive of Aspergillus spp. Aspergillus niger was isolated by microbiological examination., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Crotalaria retusa L. (rattleweed), estimated to contain about 4.96% monocrotaline (MCT) in the seed, was associated with a natural poisoning outbreak in goats. The poisoning was experimentally reproduced by the administration of C. retusa seeds containing approximately 4.49% of MCT. Thus, 1 of 3 goats given a single dose of 5 g/kg bodyweight (bw) of seeds (248 mg MCT/kg bw) and 2 goats given a single dose of 347 mg MCT/kg bw showed acute clinical signs and were euthanized 10-11 days after dosing. Clinical signs and gross and histologic lesions were characteristic of acute centrilobular liver necrosis.

Clinical, histopathological and ultrastructural findings of caprine dystrophic epidermolysis bullosa (DEB) with autosomal recessive inheritance are reported. The goats presented with exungulation, erosions, crusts and scars on the skin and ulcers in the oral cavity. Microscopically, the skin showed subepidermal separation with clefts filled occasionally with clear eosinophilic fluid, cellular debris or neutrophils. Ultrastructurally, the site of blister formation was the sublamina densa in the epidermal basement membrane zone. In skin with blister formation and in clinically uninvolved skin, the basal lamina was preserved, but the anchoring fibrils were sparse and rudimentary. A twin brother of an affected kid was mated over 5 years with his mother; three out of the 10 kids born presented with epidermolysis bullosa, indicating that the disease has an autosomal recessive mode of inheritance. It is suggested that the disease is similar to human severe generalized recessive DEB., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In this study, we investigated the presence of gastrointestinal helminths in 97 captive birds (Psittaciformes and Accipitriformes) necropsied between June and December 2011 in the state of Paraiba, Brazil. Forty-three birds were infected. Psittaciformes were infected by nematode Ascaridia hermaphrodita (97.6%) and cestode Raillietina sp. (2.4%). A. hermaphrodita was found in all species of parrots and Raillietina sp. was found only in Amazona aestiva. A. hermaphrodita was the cause of death, by intestinal obstruction, in 14 of the 40 birds investigated. Accipitriformes were infected by nematode Synhimantus (Synhimantus) rectus (100%) and acantocephalan Centrorhynchus tumidulus (50%). In Brazil, Diopsittaca nobilis and A. aestiva are reported for the first time as hosts of A. hermaphrodita and Raillietina sp., respectively. We concluded that Psittaciformes and Accipitriformes in captivity are affected by nematodes, cestodes and acanthocephalans and that implementation of control measures is essential.

Studies on the parasitic fauna of migratory sea birds of the Puffinus genus are scarce. Therefore, the aim of this study was to identify parasites of 16 specimens of Puffinus puffinus (Procellariiformes, Procellariidae) that died during the period of June 2011 to December 2011 at the Wildlife Screening Center (CETAS) of the Brazilian Institute of Environment and Natural Resources (IBAMA) in Cabedelo, Paraíba. During necropsy, biting lice and/or gastrointestinal helminths were collected in seven (43.7 %) birds. Lice were collected in five (31.2 %) birds, and the species identified were Halipeurus diversus, Trabeculus aviator, Austromenopon paululum), Saemundssonia sp. and Naubates sp. The prevalence of helminths was also 31.2 %. The nematodes species were Seuratia shipleyi and Contracaecum sp., and cestodes were Tetrabothrius sp. This is the first record in Brazil of Naubates sp., Seuratia shipleyi, Contracaecum sp., and Tetrabothrius sp. in Puffinus puffinus.

Platynosomum fastosum is a feline biliary tract trematode that generally causes asymptomatic infections. In the early 1980s in Brazil, P. fastosum was associated with cholangiocarcinomas, but this finding was not confirmed in the various publications on the parasite during the last 30 years. This study aims to report three cases of cholangiocarcinomas in cats associated with the presence of P. fastosum. From 2000 to 2011, in the Veterinary Hospital of the Federal University of Campina Grande in northeast Brazil, 348 cats were necropsied, 11 of which (3.16%) were parasitized by P. fastosum. Three cases that resulted in death were associated with cholangiocarcinomas that were found to be associated with P. fastosum. Histologically, the tumors consisted of acini composed of cells with pleomorphic nuclei, loose chromatin, evident nucleoli and lightly eosinophilic cytoplasm. Metastases were observed in two cases. The first case involved metastases to the lungs, kidneys, ovary and peritoneum, and in the second case, the lymph nodes, kidneys, heart and encephalon were involved. The other 8 cats died from other causes, and the parasite was an incidental finding. In those cases, the histologic lesions were nonsuppurative cholangiohepatitis and periductal fibrosis with P. fastosum present. Six animals also showed pre-neoplastic changes (hyperplasia and dysplasia) of the biliary duct epithelium. The study concluded that, as observed in other human biliary tract trematodes, P. fastosum causes cholangiocarcinomas in the liver of cats., (Copyright © 2012. Published by Elsevier B.V.)

Cutaneous and rhinofacial infections by Pythium insidiosum have previously been reported in sheep in Brazil. In the current study, a new form of pythiosis involving the alimentary tract of 2 nursing lambs from 2 different farms in the semiarid region of Brazil is described. The first lamb showed food regurgitation, lethargy, and anorexia, and died 5 days after the presentation of clinical signs. The second lamb had no history of gastrointestinal disease before death. Necropsy findings were similar in both lambs. The mucosa of the esophagus, reticulum, rumen, omasum, and abomasum showed ulcerated areas covered by yellowish caseous granular exudate. The lesions were transmural and extended to the serosal surfaces, and adhesions were observed between the serosa of the forestomachs and abomasum to the liver and diaphragm. Histologic lesions consisted of pyogranulomatous necrotizing transmural esophagitis, rumenitis, reticulitis, omasitis, and abomasitis with vascular thrombosis and intralesional hyphae. Pythium insidiosum was confirmed as the etiology by immunohistochemistry and culture. The presence of sheep in the vicinity of water ponds during the hot, dry season when forage is not available in the pastures seems to be the main predisposing factor for the occurrence of pythiosis in sheep in the Brazilian semiarid region.

Primary amoebic meningoencephalitis is an acute disease of the central nervous system of humans and animals caused by Naegleria fowleri. This report describes a case of meningoencephalitis caused by N. fowleri in a crossbred, one-year-old bovine with progressive neurological signs. At necropsy there was thickening of the meninges and multifocal areas of malacia in the thalamus, caudal and rostral colliculi, parietal and occipital cortex, and cerebellum. Histologically there was multifocal necrosupurative meningoencephalitis associated with areas of malacia containing amoebic trophozoites. Immunohistochemistry of the brain was positive for N. fowleri. The disease should be included among the differential diagnosis of diseases of the central nervous system in cattle in areas where animals are exposed to hot, stagnant water., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Human poisoning by Jatropha species and poisoning when livestock have been fed processed plant material has been described. Additionally, poisoning has been experimentally reproduced in various animal models. But, no cases of poisoning in livestock grazing standing and unprocessed Jatropha spp. has been reported. This study reports the poisoning of goats with Jatropha ribifolia in the semiarid region of northeastern Brazil during the dry season. The mortality of the goats ranged from 6% to 40%. The main clinical signs were apathy, anorexia, soft feces, weight loss, and severe dehydration. The skin, lips, horns, and teeth of the affected goats were stained with a reddish pigment that is present in the J. ribifolia plant. Emaciation was the main lesion observed in one necropsied goat. In 2 out of 3 goats that ingested a single dose of J. ribifolia, 10 g or 20 g of leaves of the plant per kg body weight (g/kg), mild dehydration and soft feces were observed. The plant was also administered daily to two goats for 8 days. One animal received 10 g/kg per day, and the other received 20 g/kg per day and the goats showed clinical signs after 4 and 3 days, respectively. The goat that received 10 g/kg daily recovered, and the other was euthanized. The clinical signs and lesions that were observed were similar to those observed in the spontaneous cases. This is the first case of Jatropha spp. poisoning in grazing animals that ingested the plant spontaneously., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

To determine the teratogenic effect of Mimosa tenuiflora, the green fresh plant was administered ad libitum to 12 goats (Group 1) from day 1 to day 30 of gestation. Upon ultrasonographic examination, on day 30, not one of these goats was pregnant, demonstrating that M. tenuiflora causes embryonic death. Six goats (Group 2) ingested M. tenuiflora from day 30 to day 60 of pregnancy. Four goats delivered seven healthy kids and two were not pregnant based on ultrasonographic examination on day 45 suggesting late embryonic death. Three other groups of six goats each received the plant on days 60-90 (Group 3), 90-120 (Group 4), and 120-150 (Group 5) of gestation and a control group (Group 6) all delivered normal kids, except one goat in Group 4 that aborted and one adult goat from Group 5 that was found dead. It is concluded that M. tenuiflora causes embryonic death. The failure to induce malformations might have resulted from a high dose of an unknown active principle of the plant causing fetal death., (Copyright © 2012. Published by Elsevier Ltd.)

A case of epidermolysis bullosa in a calf descendent from a Gir bull and a Gir crossbreed cow is reported. The calf presented with exungulation of all hooves, widespread erosions and crusts on the skin, and ulcers in the oral cavity. Histologically, the skin showed subepidermal separation with clefts occasionally filled with eosinophilic clear fluid, cellular debris, or neutrophils. Ultrastructurally, there was epidermal-dermal separation at the level of the lamina lucida, with the lamina densa attached to the papillary dermis. The hemidesmosomes were poorly defined and small. The clinical, histological, and ultrastructural findings are characteristic of junctional epidermolysis bullosa.

The aim of this research was to study the contribution of Toxoplasma gondii to reproductive failure using nested PCR and histopathological examination of fetuses, stillborns and placentas. We examined 245 organs of fetuses and 28 placentas from 35 abortions and stillborns from naturally occurring miscarriages in sheep in the State of Pernambuco, Brazil. At necropsy, fragments of brain, cerebellum, medulla, lung, heart, spleen, liver and placenta were taken for nested PCR and histopathological tests. Pathological examination revealed macroscopic lesions, suggesting T. gondii infection in 5/35 (14.3%) of the placentas. The histopathological examination revealed no lesions characteristic of toxoplasmosis in the organs investigated. In the five placentas, lesions consistent with toxoplasmosis were observed as an inflammatory non-suppurative infiltrate, along with multiple necrosis and mineralization. Nested PCR showed three aborted fetuses and two stillborns (14.3%) to test positive for T. gondii, with DNA amplification in all organs and the placenta, especially the heart and the placenta, which are the tissues of choice. This study substantiates the theory that T. gondii is involved in miscarriages and stillbirths and in the placentas of naturally infected sheep in Brazil. Such findings have not previously been described in the national literature., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Here, we report an outbreak of Trypanosoma vivax-induced trypanosomosis in Brazilian hair sheep on a farm in Paraíba state, a non-endemic region in northeastern Brazilian. Of 306 total sheep, 240 showed clinical signs and 216 died. Clinical signs included anorexia, lethargy, anemia, rough hair coat, weight loss, submandibular edema, abortion, and in some cases, neurological signs such as head pressing, lateral recumbence, paddling movements and muscle tremors. T. vivax was identified by blood smear analysis and polymerase chain reaction (PCR). At necropsy, animals exhibited watery blood, pale tissue coloring, and the presence of liquid in the peritoneal cavity and pericardial sac. Histologically, nonsuppurative myocarditis and meningoencephalitis with areas of malacia were observed. After treatment, no parasites were detected by blood smear analysis or PCR. Cattle and buffalo that remained in the same pasture were also infected but presented with asymptomatic infections. Epidemiological data suggest that T. vivax was introduced to the farm and the susceptible flock by buffalos that were asymptomatic carriers of the infection; T. vivax was most likely transmitted by Tabanus spp. bites and also iatrogenically., (Copyright © 2011 Elsevier B.V. All rights reserved.)

After the diagnosis of acute Crotalaria retusa poisoning, 21 healthy sheep from the affected flock were returned to the paddock and continued to consume the sprouting plants. Two years after returning these sheep to the paddock, very few plants were still alive, and after 3 years, no C. retusa plants were observed. The sheep had neither clinical signs nor biochemical alterations and delivered healthy lambs. It is concluded that resistant sheep can be used for the biological control of C. retusa., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Cryptococcal meningitis and encephalitis is reported in a 5-year-old bull. The clinical signs included multifocal neurological deficits manifested by hypermetria, ataxia, depression, circling, impaired vision, head pressing, low head carriage, wide-based stance, and falling to the side or backwards. At necropsy, 6 cavities with a gelatinous content were observed in the brain and brain stem, and the meninges were thickened and off white. Numerous yeasts, with a nonstaining capsule, and associated mild inflammatory reaction were observed histologically. Specimens for culture were not collected at necropsy, but the identity of the yeast in tissue sections as Cryptococcus neoformans var. grubii was supported by immunohistochemistry.

Outbreaks of eastern equine encephalitis observed from May 2008 to August 2009 in the Brazilian states of Pernambuco, Ceará, and Paraíba are reported. The disease occurred in 93 farms affecting 229 equids with a case fatality rate of 72.92%. Main clinical signs were circling, depression or hyperexcitability, ataxia, and progressive paralysis with a clinical manifestation period of 3-15 days. Main histologic lesions were a diffuse lymphocytic encephalomyelitis with neuronal death, satellitosis, neuronophagia, and hemorrhages being more severe in the cerebral gray matter of the telencephalon, diencephalon, and mesencephalon. Some animals also had areas of malacia in the telencephalon, thalamus, and basal nuclei. From 1 case, the virus was isolated by mice inoculation, and in other 13 cases was identified as Eastern equine encephalitis virus by semi-nested reverse transcription polymerase chain reaction. After DNA sequencing, all samples were identified as eastern equine encephalitis through the BLASTn analysis, but samples from the Ceará and Paraíba states corresponded to the same cluster, while the sample from the state of Pernambuco corresponded to a different cluster., (© 2011 The Author(s))

Sodium metabisulfite (SMB, Na(2)S(2)O(5)) is widely used in the food and pharmaceutical industries, because of its ability to inhibit proliferation of microorganisms and its antioxidant properties. We have evaluated the genotoxic effects of SMB on different tissues of the mouse, by use of the comet assay (liver and blood cells) and the micronucleus test (blood and bone marrow cells). For all tissues, significant increases in damage index and damage frequency values were observed in the SMB-treated groups (1 and 2g/kg doses) compared to the control animals. The Kruskal-Wallis test showed that the mean micronucleus frequencies in peripheral blood and bone marrow cells of mice treated with the highest dose of SMB (2g/kg) showed significant increases, when compared with controls, and a significant reduction in the ratio of polychromatic to normochromatic erythrocytes was also seen. No difference in results between sexes was observed. Our results show that high oral doses of SMB may pose a genotoxic risk., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Sodium metabisulfite is used in marine shrimp harvesting to prevent the occurrence of black spots. Shrimps are soaked in a sodium metabisulfite solution in ice, which is disposed of in sewages that run into marine canals, creating an environmental hazard. This study evaluates the toxicity and mutagenicity caused by sodium metabisulfite in sea waters and sediments collected in a shrimp farm in Cajueiro da Praia (Luis Correia), state of Piauí, Brazil, using the Allium cepa assay. Water and sediment samples were collected in the dry and in the rainy seasons, in three sites: upstream the shrimp farm (Site 1), at the point sodium metabisulfite is discharged (Site 2), and 100 m downstream the farm (Site 3). Three sample dilutions were used (50%, 25% and 10%) for all samples. A negative control (well water) and a positive control (copper sulfate 0.0006 mg mL⁻¹) were used in each experiment. At the end of the 72-h exposure period, onion roots were measured and removed. Mutagenicity analysis included the determination of mitotic index, chromosomal aberrations and the detection of micronuclei; analysis of root size and mitotic index were used as an index of toxicity. The A. cepa assay revealed that the water and sediments samples collected in the Piauí coast contaminated with sodium metabisulfite induce toxicity. The results demonstrate that the assay may be used as a regular tool in the analysis of water parameters in shrimp farms in the coast of Piauí state, and in strategies to preserve the region's marine ecosystem., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Focal symmetrical encephalomalacia (FSE) is the most prominent lesion seen in the chronic form of enterotoxemia caused by Clostridium perfringens type D in sheep. However, this lesion has not been reported in goats. The current paper reports a case of FSE in a goat from the state of Paraíba in the Brazilian semiarid region. As reported by the farmer, 30, 4-48-month-old animals from a flock of 150 goats died after showing nervous signs, including blindness and recumbence, for periods varying between 1 and 14 days. The flock was grazing native pasture supplemented with wheat and corn bran. Additionally, lactating goats were supplemented with soybeans. A 4-month-old goat with nervous signs was examined clinically and then necropsied 3 days after the onset of clinical signs. Bilateral, focal, and symmetrical areas of brown discoloration were observed in the internal capsule and thalamus. Histologic lesions in these areas consisted of multifocal, bilateral malacia with a few neutrophils; endothelial cell swelling; perivascular edema; and hemorrhages. The etiology of these lesions was not determined. However, FSE is considered pathognomonic for C. perfringens type D enterotoxemia in sheep, and it is speculated that this microorganism was the etiologic agent in the present case. The flock had been vaccinated against type D enterotoxemia only once, approximately 3 months before the beginning of the outbreak. Insufficient immunity due to the incorrect vaccination protocol, low efficacy of the vaccine used, and a diet including large amounts of highly fermentable carbohydrates were suspected to be predisposing factors for this outbreak.

Seeds of Crotalaria retusa containing 6.84% (dry weight) of monocrotaline (MCT) were administered experimentally to sheep. Three sheep that received 136.8mg MCT/kg bw daily for 70 days had no clinical signs. Five out of six sheep ingesting single doses of 205.2 and 273.6mg MCT/kg bw died with acute (three sheep) or chronic intoxication (two sheep). Acute intoxicated sheep had periacinar liver necrosis and chronic intoxicated sheep liver fibrosis and megalocytosis. Another three sheep had no clinical signs after the ingestion of 20 daily doses of 136.8mg MCT/kg, followed by seven doses of 273.6mg MCT/kg, and one single dose of 342mg MCT/kg. These experiments demonstrated that sheep are susceptible to acute intoxication by MCT being intoxicated by a single oral dose of approximately 205.2mg/kg. In contrast, they develop strong resistance to MCT after the daily ingestion of non lethal doses (136.8mg/kg). It is suggested that chronic poisoning does not occur by the repeated ingestion of non acutely toxic doses, but probably by the ingestion of single toxic doses. It is also suggested that sheep do not become intoxicated with the ingestion of C. retusa in the vegetative non-seeding stage.

A disease causing anorexia and ruminal indigestion in cattle and goats, and also edema of the lips, tongue and face in goats, was associated with the ingestion of Centratherum brachylepis in pastures containing large amounts of the plant. On 3 farms with a total of 217 cattle and 140 goats, 57 (26%) cattle and 56 (40%) goats were affected, and 11 (5%) cattle and 34 (24%) goats died. In one cow that died there were widespread and severe histologic lesions in the rumen that consisted of vacuolation and ballooning degeneration of keratinocytes, and vesicle and pustule formation in the epithelium. C. brachylepis was administered orally to 3 sheep and 11 goats. Clinical signs similar to those observed in spontaneous field cases in goats were reproduced in 2 sheep and 3 goats that ingested 30-50 g/kg body weight of the plant when administered within 48 h of it being collected. C. brachylepis collected between 2 and 13 days prior to being administered caused no clinical signs in 1 sheep and 8 goats at dose rates of 30-300 g/kg body weight of the plant. These feeding studies provide evidence that C. brachylepis is the cause of the field disease observed and that the plant loses toxicity after harvesting.

The present paper proposes a method for molecular spectrophotometric determination of copper in sugar cane spirits. The copper(I) reacts with biquinoline forming a pink complex with maximum absorption at 545 nm. The reaction occurs in the presence of hydroxylamine, ethanol and Triton X-100 tensioative. Determination of copper is possible in a linear range 0.2-20.0 mgL(-1) with a detection limit 0.05 mgL(-1). The great advantages of the proposed methodology are the elimination of liquid-liquid extraction step and the use of toxic organics solvents, like dioxane, to dissolve the reagent.

A case of protothecosis is reported in an adult goat with inspiratory dyspnea and stertor. Dermatitis with prominent ulcerated nodules up to 3 cm in diameter was observed in the muzzle at the mucocutaneous junctions of nasal and lip skin, and in the border of the pinna. Histologic lesions were necrotizing pyogranulomatous dermatitis and rhinitis with myriads of walled sporangia, characteristic of Prototheca wickerhamii. This seems to be the first report of protothecosis in a goat.

A disease known as water belly (barriga d'água), characterized by chronic progressive ascites, affects sheep in the semiarid region of northeastern Brazil. The objectives of this investigation were to study the disease and to determine its cause. Only sheep grazing for long periods in pastures where Tephrosia cinerea represents 80% to 100% of the available forage are affected. Most animals die after a clinical manifestation period of some weeks or months, but others recover when they are withdrawn from the pastures or after the first rains. At necropsy, large amounts of liquid were found in the abdominal cavity, and the liver was hard, with an irregular surface. On histology examination, the main liver lesion was chronic periportal and subcapsular fibrosis with bridging. The disease was produced experimentally in 1 sheep by the administration of large amounts of T. cinerea for 232 days. Another sheep, recovered from the spontaneous disease, had clinical signs after the ingestion of large amounts of the plant for 40 days. Seeds and leaves of the plant were examined by gas chromatography-mass spectrometry for the presence of pyrrolizidine alkaloids with negative results. It is concluded that the disease is caused by the ingestion of T. cinerea.

Craniofacial anomalies, eye malformations, and permanent flexures of the forelimbs are common malformations seen in ruminants grazing semiarid rangelands of Northeastern Brazil. To investigate the cause of these malformations, we fed 2 suspected plants, Mimosa tenuiflora or Prosopis juliflora, to groups of 4 pregnant goats each. Fresh green M. tenuiflora was collected daily and fed ad libitum to 4 goats in group 1 throughout pregnancy. This treatment group also received a supplemental feed concentrate equivalent to 1% body weight. Four goats in group 2 received a ration with 70% of P. juliflora pods and 30% hay throughout pregnancy. Four control goats were fed supplemental feed concentrate (1% body weight) and hay ad libitum throughout pregnancy. Goats treated with P. juliflora pods and the control goats delivered 9 normal kids. The four goats that were fed M. tenuiflora during pregnancy delivered 4 kids, 3 of which had abnormalities similar to those observed in field cases, including cleft lip, unilateral corneal opacity, ocular bilateral dermoids, buphthalmos with a cloudy brownish appearance of the anterior chamber due to an iridal cyst, and segmental stenosis of the colon. Malformations induced experimentally by M. tenuiflora were similar to those observed in field cases, suggesting that M. tenuiflora is a cause of the field cases observed in the Brazilian semiarid rangelands.

A disease of the central nervous system in goats was observed in the municipalities of Juazeiro, Casa Nova and Curaça, state of Bahia, and Petrolina, state of Pernambuco, Northeastern Brazil. The disease was produced experimentally in two goats by the administration of dry Turbina cordata mixed with grain. Clinical signs were observed after the ingestion of 62 and 106 g/kg body weight in 28 and 54 days, respectively. The concentration of swainsonine in the plant varied from less than 0.001% to 0.14% (dry weight). Clinical signs of natural and experimental cases included difficulties in standing, ataxia, hypermetria, wide-based stance, intention tremors, spastic paresis mainly in the hind legs, nystagmus, abnormal postural reactions, head tilting, and falling. Diffuse vacuolation of neurons, epithelial cells of pancreas, thyroids, and renal tubules were observed on the histology. From the electron microscopy of Purkinje cells the vacuoles represented dilated lysosomes. These findings demonstrated that T. cordata causes an acquired glycoprotein lysosomal storage disease. The intoxication occurs at least in an area of 27,000 km2 causing severe losses in goats, and some farmers report the disease also in cattle.

Acute intoxication by seeding Crotalaria retusa occurred during the dry season, in a flock of 80 sheep, in the semi-arid region of the state of Paraiba, northeastern Brazil. Anorexia, severe depression, mild jaundice, incoordination and recumbence were observed in 16 sheep that died within 12 h. At necropsy the liver had a nutmeg appearance. Seeds of C. retusa were found in large amounts in the rumen of the dead animals. Histologic lesions of the liver were characterized by centrilobular necrosis. Seeds of C. retusa were given to six sheep at doses of 2.5 (two sheep), 5, 10, 20 and 40 g/kg bw (one sheep for each dose). Sheep ingesting 5-40 g/kg bw were killed when moribund, 38-120 h after the start of the administration. Clinical signs and gross and histologic lesions were similar to those observed in field outbreak. The seeds used in the experiment contained 1.4% of monocrotaline. It is concluded that hungry sheep can be affected spontaneously by acute monocrotaline intoxication when they ingest large amounts of C. retusa seeds in a short period of time.

In this work, a solid phase spectrophotometric method in association with flow injection analysis for formaldehyde determination has been developed with direct measurement of light-absorption in C(18) material. The 3,5-diacetyl-1,4-dihydrolutidine produced from the reaction between formaldehyde and fluoral P was quantitatively retained on C(18) support and the spectrophotometric detection was performed simultaneously at 412nm. The retained complex was quickly eluted from C(18) material with the eluent stream consisting of a 50% (v/v) ethanol solution. The results showed that the proposed method is simple, rapid and the analytical response is linear in the concentration range of 0.050-1.5mgL(-1). The limit of detection was estimated as 30mugL(-1) and the R.S.D. 2.2% using a sample volume of 625muL. The system presented an analytical throughput of 20 determinations per hour. The method was successfully applied in the determination of formaldehyde in ethanol fuel.

Three outbreaks of hepatogenous photosensitization caused by pods of Enterolobium contortisiliquum are reported in cattle. Clinical signs were anorexia, depression, photosensitization and abortion. Most affected cattle recovered in 30-40 d. At necropsies, liver was present, the gallbladder was enlarged and edematous, and numerous seeds of E contortisiliquum were in the forestomachs and abomasum. Fruits of the plants administrated to 2 calves produced clinical signs and 2/4 died. Clinical chemistry, gross necropsies and histopathology confirmed gastrointestinal irritation and liver degeneration. One calf dosed with only E contortisiliquum leaves did not develop clinical signs.

A procedure for separation and preconcentration of trace amounts of copper in natural water samples, has been proposed. It is based on the adsorption of copper(II) ions onto a column of Amberlite XAD-2 resin loaded with calmagite reagent. This way amounts of copper within the range from 0.0125 to 25.0 microg, in a sample volume of 25 to 250 ml, and pH from 3.7 to 10.0 was concentrated as calmagite complex in a column of 0.50 g of Amberlite XAD-2 resin. Copper (II) ion was desorpted by using 5.0 ml of 2 mol l(-1) hydrochloric acid. Detection and determination limits of the proposed procedure for 250 ml sample volume were 0.15 and 0.50 microg l(-1), respectively. Selectivity test showed that (in the indicated concentration), calcium(II) (500 mg l(-1)), magnesium(II) (500 mg l(-1)), strontium(II) (50 mg l(-1)), iron(III) (10 mg l(-1)), nickel(II) (10 mg l(-1)), cobalt(II) (10 mg l(-1)), cadmium(II) (10 mg l(-1)) and lead(II) (10 mg l(-1)) did not interfere in copper determination by this procedure. Precision of the method, evaluated as the relative standard deviation by analyzing a series of seven replicates, was 2.42% for a copper mass of 1.0 microg in a sample volume of 100 ml. The accuracy of the proposed procedure was evaluated by means of copper determination in reference biological samples. The achieved results were in good agreement with certified values. The extractor system had a sorption capacity of 1.59 mumol of copper per gram of resin loaded with calmagite. The proposed procedure was applied for copper determination by FAAS in natural water samples. Samples were collected from different places of Salvador city, Bahia, Brazil. The achieved recovery, measured by the standard addition technique, showed that the proposed procedure had good accuracy. A good enrichment factor (50x) and simplicity are the main advantages in this analytical procedure.

In the present paper, a solid phase extraction system for separation and preconcentration of nickel (ng g(-1)) in saline matrices is proposed. It is based on the adsorption of nickel(II) ions onto an Amberlite XAD-2 resin loaded with 1-(2-pyridylazo)-2-naphthol (PAN) reagent. Parameters such as the pH effect on the nickel extraction, the effect of flow rate and sample volume on the extraction, the sorption capacity of the loaded resin, the nickel desorption from the resin and the analytical characteristics of the procedure were studied. The results demonstrate that nickel(II) ions, in the concentration range 0.10-275 microg l(-1), and pH 6.0-11.5, contained in a sample volume of 25-250 ml, can be extracted by using 1 g Amberlite XAD-2 resin loaded with PAN reagent. The adsorbed nickel was eluted from the resin by using 5 ml 1 M hydrochloric acid solution. The extractor system has a sorption capacity of 1.87 mumol nickel per g of Amberlite XAD-2 resin loaded with PAN. The precision of the method, evaluated as the R.S.D. obtained after analyzing a series of seven replicates, was 3.9% for nickel in a concentration of 0.20 microg ml(-1). The proposed procedure was used for nickel determination in alkaline salts of analytical grade and table salt, using an inductively coupled plasma atomic emission spectroscopy technique (ICP-AES). The standard addition technique was used and the recoveries obtained revealed that the proposed procedure shows good accuracy.

Two outbreaks of Amaranthus retroflexus (redroot pigweed) poisoning occurring in cattle in southern Brazil in late summer and early autumn are described. In both instances too many cattle were held in small paddocks heavily invaded by A retroflexus in its seeding stage. In 1 herd 8/28 heifers and in the other 15/45 adult cows died. Clinical courses ranged from 3 to 7 d. Clinical signs included depression, loss of weight, mild serous nasal discharge, foul smelling liquid feces tinged with blood, subcutaneous dependent edema, and laborious and incoordinated walking. Main necropsy findings were subcutaneous, cavitary and perirenal edemas, renal subcapsular hematomas, and ulcerative lesions in the alimentary tract. The kidneys were swollen and pale. Histopathological findings were in the kidneys and consisted of tubular degeneration, necrosis and regeneration with interstitial fibrosis and tubular proteinosis.

The reaction of aluminium(III) with Hydroxynaphtol Blue (HNB) in aqueous media at apparent pH 5.5 results in a red complex that is stable for at least 4 hr. Beer's Law is obeyed up to 1.6 microg/ml of aluminium(III) with an apparent molar absorptivity of 1.66 x 10(4) l.mol(-1). cm(-1) at 569 nm. This paper proposes procedures for aluminium(III) determination by ordinary and first-derivative spectrophotometry. The results demonstrated that the linear dynamic range is 0.03-1.60 microg/ml for ordinary spectrophotometry and 11.8-320.0 ng/ml for first derivative spectrophotometry. The HNB is not selectivity for aluminium, but the addition of EDTA allows the aluminium determination in the presence of accepted amounts of Ca(II), Mg(II), Mn(II), Ba(II), Sr(II), Cd(II), Pb(II), La(III), In(III), Bi(III) and Zn(II). The interference of Cu(II) and Hg(II) can be masked by thiosulphate. Ions such as UO(2)(II), Mo(VI), Co(II), Ti(IV) and PO(4)(III) do interfere seriously. This method was applied for aluminium determination in copper-base alloy, zinc-base alloy, magnesium-base alloy, iron ore, manganese ore, cement, dolomite, feldspar and limestone. The results indicated high accuracy and precision.