Physiological response, function of sweat glands, and hair follicle cycling in cattle in response to fescue toxicosis and hair genotype.

Fescue toxicosis is a syndrome that results when cattle consume toxic endophyte-infected tall fescue. The objective of this study was to compare the response in physiological variables, sweat gland function, hair follicle cycling, and gene expression to feeding a total mixed ration that included tall fescue haylage and tall fescue seed containing a toxic endophyte (EI) or tall fescue haylage containing a nontoxic novel endophyte (EN) in beef heifers (Angus × Senepol heifers, n = 31) with 2 different hair genotypes. Numbers in each subgroup were as follows: novel endophyte, heterozygous slick (EN-S; n = 8), novel endophyte, homozygous hairy (wild type, EN-W; n = 7), endophyte-infected, heterozygous slick (EI-S; n = 10), and endophyte-infected, homozygous hairy (wild type, EI-W; n = 6). Physiological measurements were taken weekly for 7 wk. Data were analyzed using the MIXED procedure of SAS including dietary fescue treatment (EN vs. EI) and hair genotype (S vs. W) as main effects, day as a repeated measure, and temperature-humidity index (THI) as a covariate. Skin biopsies were taken before treatment initiation and on day 37 of treatment. Average surface temperature (ST) increased as the THI increased (P < 0.0001). Average ST was greater (P < 0.01) for animals fed EI than for animals fed the EN fescue diet, and greater (P < 0.01) for animals with the W genotype compared with animals with the S genotype. The difference between heifers with the S and W genotype was greater at greater THI (genotype × day interaction, P < 0.01). Transepidermal water loss (TEWL) was greater (P < 0.05) for animals with the S genotype compared with the W genotype and greater (P < 0.05) for heifers with the S genotype than for heifers with the W genotype when fed EI (36.7, 38.5, 30.0, and 38.7 g/m2 per hour for EN-W, EN-S, EI-W, and EI-S, respectively). The fraction of follicles in telogen in plucked hair samples for heifers fed EI was greater for animals with the S genotype than the W genotype (fraction in telogen: 0.456, 0.565, 0.297, 0.702 for EN-W, EN-S, EI-W, and EI-S, respectively; diet × genotype interaction, P < 0.05). Fraction of follicles in anagen was the opposite. EI fescue resulted in increased ST, changes in hair follicle cycling that support greater hair growth, and decreased TEWL for heifers with the W genotype compared with S genotype, suggesting greater heat stress in response to EI.

Fusarium species isolated from Pennisetum clandestinum collected during outbreaks of kikuyu poisoning in cattle in South Africa.

Kikuyu poisoning occurs sporadically in South Africa. It is of major economic importance, as valuable dairy cows are often poisoned by it, and once affected, the mortality rate is high. Pennisetum clandestinum samples were collected during eight outbreaks of kikuyu poisoning in cattle in the Eastern Cape Province of South Africa from 2008 to 2010. The kikuyu grass samples were submitted specifically for the isolation and molecular identification of Fusarium species, as it was recently suggested that mycotoxins synthesised by Fusarium torulosum could be the cause of this intoxication. Ninety-four Fusarium isolates were retrieved from the grass samples, of which 72 were members of the Fusarium incarnatum/Fusarium equiseti species complex based on morphology and phylogenetic analyses of the translation elongation factor 1α sequence data. The South African isolates from kikuyu identified as members of the F. incarnatum/F. equiseti species complex grouped together in six separate clades. The other isolates were Fusarium culmorum (n = 3), Fusarium redolens (n = 4) and Fusarium oxysporum (n = 15). Although F. torulosum could not be isolated from P. clandestinum collected during kikuyu poisoning outbreaks in South Africa, the mycotoxicosis theory is still highly plausible.

Identification of protoxins and a microbial basis for red maple (Acer rubrum) toxicosis in equines.

The leaves of Acer rubrum (red maple), especially when wilted in the fall, cause severe oxidative damage to equine erythrocytes, leading to potentially fatal methemoglobinemia and hemolytic anemia. Gallic acid and tannins from A. rubrum leaves have been implicated as the toxic compounds responsible for red maple toxicosis, but the mechanism of action and toxic principle(s) have not been elucidated to date. In order to investigate further how red maple toxicosis occurs, aqueous solutions of gallic acid, tannic acid, and ground dried A. rubrum leaves were incubated with contents of equine ileum, jejunum, cecum, colon, and liver, and then analyzed for the metabolite pyrogallol, as pyrogallol is a more potent oxidizing agent. Gallic acid was observed to be metabolized to pyrogallol maximally in equine ileum contents in the first 24 hr. Incubation of tannic acid and A. rubrum leaves, individually with ileum contents, produced gallic acid and, subsequently, pyrogallol. Ileum suspensions, when passed through a filter to exclude microbes but not enzymes, formed no pyrogallol, suggesting a microbial basis to the pathway. Bacteria isolated from ileum capable of pyrogallol formation were identified as Klebsiella pneumoniae and Enterobacter cloacae. Therefore, gallotannins and free gallic acid are present in A. rubrum leaves and can be metabolized by K. pneumoniae and E. cloacae found in the equine ileum to form pyrogallol either directly or through a gallic acid intermediate (gallotannins). Identification of these compounds and their physiological effects is necessary for the development of effective treatments for red maple toxicosis in equines.

Intoxicação por Brachiaria spp. em bovinos no Mato Grosso do Sul / Brachiaria spp. poisoning in cattle in Mato Grosso do Sul, Brazil

Mediante a revisão dos arquivos das fichas de necropsia do Laboratório de Patologia Animal, Universidade Federal do Mato Grosso do Sul, foram estudados a epidemiologia, o quadro clínico e a patologia de 29 surtos de intoxicação por Brachiaria spp., ocorridos em bovinos de corte, no Mato Grosso do Sul, de março de 1996 a novembro de 2009. Os surtos ocorreram em todas as épocas do ano, tanto na seca quanto na chuva. Em 24 dos 29 surtos o principal sinal clínico foi a fotossensibilização e em cinco o principal sinal foi o emagrecimento progressivo. Dos 24 surtos de fotossensibilização, 11 ocorreram em pastagens de B. decumbens, dois em pastagens mistas de B. decumbens e B. brizantha, um em B. brizantha e em 10 surtos não foi informada a espécie de Brachiaria envolvida. A morbidade variou de 0,2 por cento a 50 por cento e a letalidade de 44,4 por cento a 100 por cento. Nos casos de fotossensibilização o edema de barbela foi o sinal clínico mais encontrado em bovinos, seguido de dermatite com pele espessada no flanco e períneo, retração cicatricial auricular, icterícia, corrimento ocular. crostas auriculares e oculares, e ulcerações na parte ventral da língua. Em dois surtos foram observados sinais nervosos e em um, diarréia. Nas necropsias o fígado estava aumentado de tamanho, amarelado, com padrão lobular aumentado e, ocasionalmente, com áreas esbranquiçadas e deprimidas. Os rins estavam acastanhados e a urina escura. No exame histológico do fígado encontrou-se tumefação e vacuolização de hepatócitos, proliferação de células epiteliais dos ductos biliares, retenção biliar, fibroplasia periportal discreta ou moderada e infiltrado mononuclear periportal. Todos os casos de fotossensibilização apresentaram macrófagos espumosos no parênquima hepático e em 21 foram observados cristais birrefringentes nos ductos biliares. Cinco surtos com emagrecimento progressivo dos bovinos afetados foram diagnosticados em pastagens de B. decumbens. A principal lesão macroscópica foi o fígado aumentado de volume e amarelado. No estudo histológico, as lesões foram semelhantes às observadas nos casos de fotossensibilização, sendo que cristais refringentes nos ductos biliares foram observados em três animais. Conclui-se que B. decumbens é mais tóxica que B. brizantha e que a intoxicação ocorre, principalmente, em bovinos jovens nas diferentes épocas do ano. No entanto, pesquisas são necessárias para determinar as variações no conteúdo de saponinas litogênicas em Brachiaria spp. e as diferenças de resistência/susceptibilidade à intoxicação de bovinos de diferentes idades e raças em diferentes regiões.

The epidemiology, clinical signs and pathology of 29 outbreaks of spontaneous poisoning by Brachiaria spp. in beef cattle, which occurred from March 1996 to November 2009, in the state of Mato Grosso do Sul, were retrospectively studied. For this, the files of the Veterinary Pathology Laboratory at the Federal University of Mato Grosso do Sul were reviewed. Outbreaks occurred at different times of the year, including dry and rainy seasons. While photosensitization was the main clinical sign in cattle from 24 out of 29 outbreaks, in five of them the main clinical sign was progressive wasting, without photosensitization. Eleven outbreaks of photosensitization occurred in pastures of Brachiaria decumbens, two in mixed pastures of B. decumbens and B. brizantha, and one in B. brizantha. In 10 outbreaks the species of Brachiaria was not informed. Morbidity ranged from 0.2 percent to 50 percent, and fatality rates were between 44.4 percent and 100 percent. In cases of photosensitization brisket edema was the most frequent clinical sign observed. Dermatitis with thickened skin of the flank and perineal region, scar retraction of the ears, jaundice, ocular discharge, crusts in the eyes and ears, and ulceration of the ventral surface of the tongue were also observed. Nervous signs were observed in two cases and diarrhea in one. At necropsy the liver was enlarged, yellowish, with increased lobular pattern, and occasionally with depressed whitish areas. The kidneys were brownish and the urine dark. Upon histological examination of the liver the hepatocytes were swollen and vacuolized. Individual necrosis of hepatocytes, bile duct cell proliferation, biliary retention, and mild to moderate periportal fibrosis were also observed. In all cases foamy macrophages, most in groups, were observed mainly in the centrilobular region. In 21 cattle birefringent crystals were observed within the bile ducts. Five outbreaks of progressive wasting were observed in cattle grazing B. decumbens pastures. The main lesion was an enlarged yellowish liver. Histologic lesions were similar to those observed in cases of photosensitization. Birefringent crystals were observed in the liver of three cases, but foamy macrophages were found in all animals. It is concluded that B. decumbens is more toxic than B. brizantha and that the poisoning occurs at any time of the year. Nevertheless, research is still necessary to determine variations in the saponin content of Brachiaria species and the probable differences on susceptibility/resistance to the poisoning of cattle of different ages, breeds and from different regions.

Intoxicação espontânea e experimental por Brachiaria radicans (tanner-grass) em bovinos / Natural and experimental poisoning by Brachiaria radicans (tanner grass) in cattle

Brachiaria radicans (tanner-grass,) cresce bem em solos úmidos. Em Santa Catarina é encontrada principalmente nas regiões dos vales dos rios Tubarão e Itajaí. Quando ingerida em grandes quantidades pelos bovinos induz anemia hemolítica, hemoglobinúria, diarreia e pode evoluir para a morte. O objetivo deste trabalho foi avaliar os dados epidemiológicos e clínico-patológicos causados por B. radicans em bovinos. A planta foi administrada a 12 bovinos em doses de 50-100 por cento da dieta. Os animais que receberam uma dieta que consistia de 100 por cento da planta, originária de solos turfosos, mostraram hemoglobinúria, diarreia, mucosas vermelho-escuras e recuperação após suspensão da ingestão da planta. Exames de sangue e urina revelaram anemia, hemoglobinúria e proteinúria. A histopatologia de material coletado de bovinos que morreram pela doença espontânea, revelou necrose hepática coagulativa e paracentral e nefrose hemoglobinúrica. B. radicans mostrou ser tóxica para bovinos somente quando cresce em solos férteis e se consistir de 100 por cento da dieta.

Brachiaria radicans is a grass that grows well on humid soils. In Santa Catarina, it is found mainly in the valleys of the Tubarão and Itajaí rivers. When eaten by cattle in large amounts induces hemolytic anemia, hemoglobinuria, diarrhea and even death. The objective of this study was to evaluate epidemiologic, clinical and pathological data of the intoxication caused by B. radicans in cattle. The plant was administered to 12 cattle in doses of 50-100 percent of the diet. The animals that received 100 percent of B. radicans, grown on peaty soils, showed hemoglobinuria, diarrhea and dark red mucous membranes; they recovered when ingestion of the grass was discontinued. Blood and urine tests revealed anemia, hemoglobinuria and proteinuria. Histopathology of organs collected from cattle that died from spontaneous poisoning revealed hepatic centrolobular and paracentralr coagulative necrosis and hemoglobinuric nephrosis. B. radicans proved toxic for cattle only when it grows on fertile soils and is ingested as 100 percent of the diet.

Annual ryegrass toxicity in Thoroughbred horses in Ceres in the Western Cape Province, South Africa.

An outbreak of annual ryegrass toxicity occurred on a Thoroughbred stud in Ceres in the Western Cape Province of South Africa. This is the 1st report of annual ryegrass toxicity in horses in South Africa, although the condition has been reported in cattle and sheep populations in the past. Annual ryegrass toxicity is characterised by a variety of neurological signs including tremors, convulsions, recumbency and in many cases death. The description of the outbreak includes the history, clinical presentation and treatment protocol administered during the outbreak. Various epidemiological variables and their influence in the outbreak are also considered.

[Young perennial ryegrass staggers in a dairy herd]. / Raaigraskramp bij jongvee op een melkveebedrijf.

Ryegrass staggers was diagnosed in a group of young stock on a dairy farm in the Netherlands. The cattle were fed on ryegrass hay originating from a farm in the region. The typical clinical signs observed in this herd were difficulty in rising, hypermetria of especially the front legs, incoordination, and tremors of the head and neck muscles. The morbidity of the illness was about 80 percent and there was no mortality. Four weeks after withdrawal of the hay, the animals recovered completely. The diagnosis was confirmed by demonstrating high concentrations of Lolitrem-B (1.7 ppm) in the hay. This neurotoxin is usually produced by the endophyte Neotyphodium lolii with which ryegrass may be infected. This is the first case of ryegrass staggers in cows reported in the Netherlands since 1992.

Two cases of equine grass sickness with evidence for soil-borne origin involving botulinum neurotoxin.

Botulism is caused by different types of Clostridium botulinum, a soil bacterium. Equine grass sickness (equine dysautonomia) is suspected of being a clinical form of this disease. On a stud where this disease occurred twice within 8 months, grass and soil samples and necropsy specimens of one horse were tested for the presence of bacterial forms and toxin of C. botulinum. Different types and type mixtures (A-E) of C. botulinum and botulinum neurotoxin were found. For the first time, it has been shown that green grass blades contain botulinum toxin. The results support the hypothesis that equine grass sickness is a clinical form of botulism, a soil-borne disease.

Detection of endophyte toxins in the imported perennial ryegrass straw.

From 1997 to 1999, 29 cases of disorders were detected in cattle and horses that had been fed ryegrass straw imported from the U.S.A. These animals showed symptoms resembling ryegrass staggers and the clinical signs disappeared after removal of the straw. Endophytic hyphae were detected in the seeds of all straw samples that were responsible for the clinical cases. Lolitrem B concentrations in the straw ranged between 972 and 3740 ppb. Ergovaline concentrations were between 355 and 1300 ppb. Even though the concentrations of lolitrem B were lower than the toxic threshold proposed by Oregon State University in better part of the cases, our observations suggest the possibility that lolitrem B lower than the proposed threshold can bring disorders to sensitive individuals.

Genetic analysis of Clavibacter toxicus, the agent of annual ryegrass toxicity.

Multilocus enzyme electrophoresis was used to examine the relatedness of 52 isolates of Clavibacter toxicus, the agent of annual ryegrass toxicity. These included 37 Western Australian (WA) field isolates sampled in 3 distinct locations over a 2-year period, and 15 isolates sampled from 6 different host plant species in 3 states in Australia over approximately 8 years. Seventeen reference strains for the related genera Curtobacterium, Rhodococcus and Arthrobacter were examined for comparison. The 69 isolates were divided into 29 electrophoretic types (ETs), separated by genetic distances of 0.06 to 0.81. The C. toxicus isolates fell into 12 ETs, 11 of which formed a tightly clustered group separated by a genetic distance of 0.23 or less. Thirty-one of the WA field isolates of C. toxicus fell into a single ET, and four into another ET. Clavibacter toxicus therefore formed a closely related group which was genetically distinct from the other plant pathogenic species, and a dominant widely disseminated strain of the species was identified in WA.

Fungi on Narthecium ossifragum leaves and their possible involvement in alveld disease of Norwegian lambs.

Spores of Pithomyces chartarum (Berk. & Curt.) M.B. Ellis were only rarely seen on leaves of Narthecium ossifragum (L.) Hudson collected in summer from five areas in western Norway in which alveld, a photosensitization disease of lambs, is endemic. Cladosporium magnusianum (Jaap) M.B. Ellis was found on all 118 leaf samples collected in the summers of 1990 and 1991. The hypothesis that P. chartarum contributes to the aetiology of alveld could not be supported, but it is possible that C. magnusianum may have a role in the causation of the disease.

Kikuyu poisoning of cattle in New South Wales and its relationship to pasture fungi on kikuyu grass (Pennisetum clandestinum).

In the late summer and autumn of 1980 kikuyu poisoning of cattle was diagnosed on 9 farms in New South Wales resulting in 213 deaths among 1370 cattle at risk. Clinical, autopsy and histopathological findings are described and are consistent with those previously recorded. Kikuyu grass samples from 5 paddocks in which deaths were occurring were examined for the presence of fungi. Myrothecium spp were not present. Fusarium semitectum, F. moniliforme var subglutinans, Penicillium spp and a Phoma sp were the most common fungi isolated but with the exception of F. semitectum were not consistently present. F. semitectum was present in adjacent kikuyu grass paddocks in which deaths were not occurring. Two of the farms had army caterpillar (Mythimna convecta) infestations. Army caterpillar faeces present in these paddocks did not contain Myrothecium spp; the fungi present reflected the population present on the kikuyu grass. These findings do not suggest a mycotic cause for kikuyu poisoning.

Effects of feeding lupin seed naturally infected with Phomopsis leptostromiformis to sheep and pigs.

Lupin seed, 10% of which was naturally infected with Phomopsis leptostromiformis was obtained from 2 different sources and stored either conventionally, or in a warm and humid environment, for 45 weeks. Only very low levels of toxicity were apparent during the storage period and in the warm, humid environment the level of infection of P. leptostromiformis was gradually reduced and eventually eliminated. The 2 batches of infected seed, and non-infected lupin seed, were fed to sheep as sole rations for 40 weeks, or as part of feedlot rations for 10 weeks, and to baconer pigs as part of grower and finisher rations for 16 weeks. In the 40-week feeding trial mild lupinosis developed in sheep fed from one batch of infected seed, but there was no evidence of lupinosis in sheep fed the feedlot rations, or pigs fed the grower and finisher rations. It is suggested that lupin seed with levels of Phomopsis-infection of up to 10% could be safely fed to sheep and pigs under commercial conditions.

Epichloë typhina from toxic tall fescue grasses.

Epichloë typhina, a clavicipitaceous systemic phytopathogen, was isolated from two varieties and three hybrids of tall fescue (Festuca arundinaceae). The morphology of the fescue isolates was compared with E. typhina isolated from bent grass (Agrostis perennans). In all isolates, conidia were identical and were typical of E. typhina. In fescue grasses the endophyte failed to produce stromata, but on bent grass the fungus seasonally produced stromata, typical of the genus. Cattle grazing the fescue grasses showed signs of the fescue toxicity syndrome, the E. typhina was found in frequencies of 100%; in grasses from pastures in which cattle showed no signs of the syndrome, frequencies were 0 to 50%. Nutritional factors in vitro were more complex for the isolates from fescue than for the isolate from bent grass. These studies suggested that E. typhina includes biotypes that might be involved in the toxicity syndrome. The fescue biotypes grew poorly on media, and yields were inadequate for toxicity studies. However, the bent grass isolate grew well on three media, and extracts from two of these were toxic to chicken embryos. All isolates produced in vitro the nontoxic fungal steroid tetraenone [ergosta-4,6,8(14),22-tetraen-3-one], which has been isolated from toxic fescue grasses.