Activation of C-C motif chemokine receptor 2 modulates testicular macrophages number, steroidogenesis, and spermatogenesis progression.

The monocyte chemoattractant protein 1 (MCP-1) belongs to the CC chemokine family and acts in the recruitment of C-C motif chemokine receptor 2 (CCR2)-positive immune cell types to inflammation sites. In testis, the MCP-1/CCR2 axis has been associated with the macrophage population's functional regulation, which presents significant functions supporting germ cell development. In this context, herein, we aimed to investigate the role of the chemokine receptor CCR2 in mice testicular environment and its impact on male sperm production. Using adult transgenic mice strain that had the CCR2 gene replaced by a red fluorescent protein gene, we showed a stage-dependent expression of CCR2 in type B spermatogonia and early primary spermatocytes. Several parameters related to sperm production were reduced in the absence of CCR2 protein, such as Sertoli cell efficiency, meiotic index, and overall yield of spermatogenesis. Daily sperm production decreased by almost 40%, and several damages in the seminiferous tubules were observed. Significant reduction in the expression of important genes related to the Sertoli cell function (Cnx43, Vim, Ocln, Spna2) and meiosis initiation (Stra8, Pcna, Prdm9, Msh5) occurred in comparison to controls. Also, the number of macrophages significantly decreased in the absence of CCR2 protein, along with a disturbance in Leydig cell steroidogenic activity. In summary, our results show that the non-activation of the MCP-1/CCR2 axis disturbs the testicular homeostasis, interfering in macrophage population, meiosis initiation, blood-testis barrier function, and androgen synthesis, leading to the malfunction of seminiferous tubules, decreased testosterone levels, defective sperm production, and lower fertility index.

Características morfológicas do estômago de jaguatirica (Leopardus pardalis - Linnaeus, 1758) / Morphological characteristics of the stomach of the ocelot (Leopardus pardalis - Linnaeus, 1758)

Leopardus pardalis é uma das espécies de felinos neotropicais mais estudadas em seus aspectos ecológicos, porém informações de questões morfofisiológicas do trato digestório não são encontradas na literatura. Visando contribuir com tais informações, o objetivo deste artigo foi caracterizar a morfologia do estômago dessa espécie. Nesta pesquisa, foram utilizados cinco exemplares adultos, coletados após óbito por atropelamento ou doados pelo IBAMA ao Laboratório de Zoologia e Morfologia Animal da Universidade do Estado de Mato Grosso - Alta Floresta. Os animais foram fixados com solução aquosa de formaldeído a 10%, dissecados por meio de técnicas anatômicas. Além dos estudos anatômicos, também foram coletados tecidos para microscopia. As análises macroscópicas revelaram que o estômago era do tipo unicavitário, com presença de pequena e grande curvatura, localizado no antímero esquerdo e constituído pelas regiões características do órgão. Em seus aspectos microscópicos, mostrou-se formado pelas quatro túnicas usuais do tubo digestório: túnica mucosa, submucosa, muscular e serosa, cada uma com as particularidades características de cada região, principalmente na túnica mucosa. O estômago de L. pardalis revelou uma morfologia semelhante às espécies domésticas e selvagens, com particularidades histomorfológicas na região pilórica. Esse padrão pode estar relacionado a adaptações evolutivas no processo digestivo dessa espécie.(AU)

The species Leopardus pardalis is one of the species of neotropical felines more studied in its ecological aspects, however, information of morphophysiological questions of the digestive tract are not found in the literature. Aiming to contribute with such information, the objective of this work was to characterize the morphology of the stomach of this species. Five adult specimens, collected after death by trampling, or donated by IBAMA to the Laboratory of Zoology and Animal Morphology of the University of the State of Mato Grosso - Alta Floresta, were used in this research. The animals were fixed with aqueous solution of formaldehyde 10%, dissected through the basic techniques and instruments of anatomy, and later the material was collected for microscopy. The macroscopic analysis revealed that the stomach was of the unicavitary type, with small and great curvature, located in the left antimer and constituted by the characteristic regions of the organ. In its microscopic aspects it was formed by the four usual tunics of the digestive tube: tunica mucosa, submucosa, muscular and serosa, each one with the peculiarities characteristic of each region, mainly in the tunica mucosa. The stomach of L. pardalis revealed similar morphology to the domestic and wild species, with small histomorphological peculiarities in the pyloric region, which may be related to evolutionary adaptations in the digestive process of this species.(AU)

Duration of spermatogenesis and identification of spermatogonial stem cell markers in a Neotropical catfish, Jundiá (Rhamdia quelen).

Spermatogenesis is a process driven by stem cell, where germ cell cycle is under the control of a specific genotype species. Considering that Jundiá (Rhamdia quelen) is a Neotropical catfish with great economical importance and useful experimental model, little information is available on basic aspects of its reproductive biology, especially on spermatogenesis. As a result, this study aimed to characterize the male germ cells, estimate the duration of spermatogenesis and evaluate the expression of selected stem cell genes in Jundiá testis. Similar to other fish species, our results showed a remarkable decrease of germ cell nuclear volume during Jundiá spermatogenesis, particularly from type A undifferentiated to late type B spermatogonia and from diplotene to late spermatids. Using a S-phase marker, bromodeoxyuridine (BrdU), the combined duration of meiotic and spermiogenic phases in this species was estimated in approximately 7 days. This is considered very short when compared to mammals, where spermatogenesis last from 30 to 74 days. Selected stem cell genes were partially sequenced and characterized in Jundiá testis. Expression analysis showed higher plzf and pou5f3 mRNA levels in the cell fractions enriched by type A undifferentiated spermatogonia. These results were further confirmed by in situ hybridization that showed strong signal of plzf and pou5f3 mRNA in type A undifferentiated spermatogonia. Altogether, these information will expand our knowledge of the reproductive biology of this species, contributing to improve its production and management, and also for biotechnological applications, such as germ cell transplantation.

Oral administration of Simbioflora® (synbiotic) attenuates intestinal damage in a mouse model of 5-fluorouracil-induced mucositis.

The use of probiotics to prevent or treat mucosal inflammation has been studied; however, the combined effect of probiotics and prebiotics is unclear. The aim of this study was to test whether oral administration of a synbiotic (Simbioflora®) preparation containing Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium lactis plus fructooligosaccharide could help control mucosal inflammation in experimental mucositis induced by 5-fluorouracil (5-FU). Male BALB/c mice were randomly divided into six groups: control (CTL), control + prebiotic (CTL+P), control + synbiotic (CTL+S), mucositis (MUC), mucositis + prebiotic (MUC+P), and mucositis + synbiotic (MUC+S). Mice from the CTL+S, MUC+S, CTL+P, and MUC+P groups received synbiotic or prebiotic daily by oral gavage for 13 days. Mice in the CTL and MUC groups received the same volume of saline. On day 11, mice in the MUC, MUC+P, and MUC+S groups received an intraperitoneal injection of 300 mg/kg 5-FU to induce mucositis. After 72 h, all mice were euthanised. Intestinal permeability, intestinal histology, and biochemical parameters were analysed. Group MUC showed a greater weight loss and increased intestinal permeability (0.020 counts per min [cpm]/g) compared to the CTL group (0.01 cpm/g) P<0.05. Both treatments attenuated weight loss compared to the MUC group. Nonetheless, the synbiotic caused a greater reduction in intestinal permeability (0.012 cpm/g) compared to the MUC (0.020 cpm/g) and MUC+P (0.016 cpm/g) groups P<0.05. Mice in groups MUC+P and MUC+S displayed significant recovery of lesions and maintenance of the mucus layer. There were no differences in the short-chain fatty acid concentrations in the faeces between the MUC and CTL groups (P>0.05). Increased acetate and propionate concentrations were evidenced in the faeces of the MUC+P and MUC+S groups. Only the synbiotic treatment increased the butyrate concentration (P<0.05). The results indicate that administration of synbiotic can decrease mucosal damage caused by mucositis.

Successful xenogeneic germ cell transplantation from Jundia catfish (Rhamdia quelen) into adult Nile tilapia (Oreochromis niloticus) testes.

Fish germ cell transplantation presents several important potential applications for aquaculture, including the preservation of germplasm from endangered fish species with high genetic and commercial values. Using this technique in studies developed in our laboratory with adult male Nile tilapias (Oreochromis niloticus), all the necessary procedures were successfully established, allowing the production of functional sperm and healthy progeny approximately 2months after allogeneic transplantation. In the present study, we evaluated the viability of the adult Nile tilapia testis to generate sperm after xenogeneic transplant of germ cells from sexually mature Jundia catfish (Rhamdia quelen) that belong to a different taxonomic order. Therefore, in order to investigate at different time-periods post-transplantation, the presence and development of donor PKH26 labeled catfish germ cells were followed in the tilapia seminiferous tubules. From 7 to 20days post-transplantation, only PKH26 labeled spermatogonia were observed, whereas spermatocytes at different stages of development were found at 70days. Germ cell transplantation success and progression of spermatogenesis were indicated by the presence of labeled PKH26 spermatids and sperm on days 90 and 120 post-transplantation, respectively. Confirming the presence of the catfish genetic material in the tilapia testis, all recipient tilapias evaluated (n=8) showed the genetic markers evaluated. Therefore, we demonstrated for the first time that the adult Nile tilapia testis offers the functional conditions for development of spermatogenesis with sperm production from a fish species belonging to a different order, which provides an important new venue for aquaculture advancement.

Viabilidade econômica da terceira ordenha em sistemas de produção de leite com ordenhadeira tipo circuito aberto / Economic viability of the third milking in milk production systems using open circuit type milking mechanics

Objetivou-se analisar a viabilidade econômica da terceira ordenha em sistemas de produção de leite que utilizam ordenhadeira mecânica, do tipo circuito aberto, visando fornecer aos técnicos e pecuaristas informações que os auxiliem no processo de tomada de decisões. Especificamente pretendeu-se ainda: a) estimar o custo de uma ordenha; b) estimar, por meio de simulação, o custo da terceira ordenha; c) desenvolver uma equação matemática que permita estimar a quantidade mínima de leite produzido em duas ordenhas, a partir da qual será viável economicamente a realização da terceira ordenha. Os dados foram coletados em três sistemas de produção de leite, em uma rotina de duas ordenhas diárias, sendo realizadas três coletas de dados em cada um, perfazendo um total de nove. Considerando-se os dados médios, seria viável a realização da terceira ordenha se a produtividade média diária das vacas em lactação em duas ordenhas fosse igual ou superior a 38,13kg de leite. A equação matemática desenvolvida pode auxiliar o técnico e o pecuarista a estimarem a quantidade mínima de leite produzido por uma vaca em duas ordenhas, a partir da qual será viável economicamente a realização da terceira ordenha, com precisão e considerável rapidez.

This study aims to analyze the economic viability of the third milking in milk production systems using open circuit milking mechanics, aiming to provide technicians and farmers with information to help in the decision-making process. Specifically intend to a) estimate the cost of a milking; (b) estimate, by means of simulation, the cost of the third milking; (c) develop a mathematical equation that allows estimating the minimum amount of milk produced in two milkings, from which it will be economically feasible to hold the third milking. The data were collected in three milk production systems, in a routine of two milkings per day, with three collections of data in each, a total of nine collections. Considering the average data, it would be feasible to hold the third milking if the productivity of daily average lactating cows is equal or greater than 38.13 kg of milk. The mathematical equation developed may help the technician and the farmer to estimate the minimum quantity of milk produced by a cow in two milkings, from which it will be economically feasible to hold the third milking with precision and considerable speed.

Germ cell transplantation as a potential biotechnological approach to fish reproduction.

Although the use of germ cell transplantation has been relatively well established in mammals, the technique has only been adapted for use in fish after entering the 2000s. During the last decade, several different approaches have been developed for germ cell transplantation in fish using recipients of various ages and life stages, such as blastula-stage embryos, newly hatched larvae and sexually mature specimens. As germ cells can develop into live organisms through maturation and fertilization processes, germ cell transplantation in fish has opened up new avenues of research in reproductive biotechnology and aquaculture. For instance, the use of xenotransplantation in fish has lead to advances in the conservation of endangered species and the production of commercially valuable fish using surrogated recipients. Further, this could also facilitate the engineering of transgenic fish. However, as is the case with mammals, knowledge regarding the basic biology and physiology of germline stem cells in fish remains incomplete, imposing a considerable limitation on the application of germ cell transplantation in fish. Furthering our understanding of germline stem cells would contribute significantly to advances regarding germ cell transplantation in fish.

Testis stereology, seminiferous epithelium cycle length, and daily sperm production in the ocelot (Leopardus pardalis).

Similar to most wild felids, the ocelot (Leopardus pardalis) is an endangered species. However, knowledge regarding reproductive biology of the ocelot is very limited. Germ cell transplantation is an effective technique for investigating spermatogenesis and stem cell biology in mammals, and the morphologic characterization of germ cells and knowledge of cycle length are potential tools for tracking the development of transplanted germ cells. Our goal was to investigate basic aspects related to testis structure, particularly spermatogenesis, in the ocelot. Four adult males were used. After unilateral orchiectomy, testis samples were routinely prepared for histologic, stereologic, and autoradiographic analyses. Testis weight and the gonadosomatic index were 11+/-0.6g and 0.16+/-0.01%, respectively, whereas the volume density of seminiferous tubules and Leydig cells was 83.2+/-1.6% and 9.8+/-1.5%. Based on the acrosomic system, eight stages of spermatogenesis were characterized, and germ cell morphology was very similar to that of domestic cats. Each spermatogenic cycle lasted 12.5+/-0.4 d, and the entire spermatogenic process lasted 56.3+/-1.9 d. Individual Leydig cell volume was 2522mum(3), whereas the number of Leydig and Sertoli cells per gram of testis was 38+/-5x10(6) and 46+/-3x10(6). Approximately 4.5 spermatids were found per Sertoli cell, whereas daily sperm production per gram of testis was 18.3+/-1x10(6), slightly higher than values reported for other felids. The knowledge obtained in this study could be very useful to the preservation of the ocelot using domestic cat testes to generate and propagate the ocelot genome.

Effects of selective serotonin reuptake inhibitor therapy on endothelial function and inflammatory markers in patients with coronary heart disease.

We investigated the effect of sertraline on inflammation and endothelial function in patients with coronary heart disease (CHD) and symptoms of depression. One hundred patients with CHD and depression were randomized in a double-blind fashion to receive sertraline or a placebo. We measured symptoms of depression (Beck Depression Inventory (BDI) score), levels of inflammatory markers (C-reactive protein (CRP) and interleukin-6 (IL-6)), and flow-dependent endothelium-mediated dilation (FMD) before and after 20 weeks of treatment. Sertraline treatment significantly reduced the BDI score as compared with both baseline and placebo. Levels of CRP and IL-6 also decreased after 20 weeks of sertraline treatment, whereas they did not significantly change in the placebo group. There was a significant improvement in FMD in patients on sertraline treatment, whereas there was no change in FMD in the placebo group. Sertraline improves endothelial function and reduces inflammatory markers in patients with CHD and symptoms of depression.

Streptococcus agalactiae associado à meningoencefalite e infecção sistêmica em tilápia-do-nilo (Oreochromis niloticus) no Brasil / Streptococcus agalactie associated to meningoencefalitis and systemic infection from tilapia (Oreochromis niloticus) in Brazil

Streptococcus agalactiae was isolated from cultured tilapia in Brasil. All isolates reacted similarly in API 20 STREP system (bioMerieux, France) and Slidex strepto grouping Latex Aglutination test (bioMerieux, France). Bacterial isolates were submitted to the disc diffusion technique for the antimicrobial susceptibility test. Among nine tested drugs S. agalactiae samples were resistant to three-neomicin, nalidixic acid and gentamicin.

Diarréia em leitões lactentes por Clostridium perfringens tipo A em granjas tecnificadas nos estados de Minas Gerais e São Paulo / Clostridium perfringens type A diarrhea in suckling piglets in industrial swine farms in the states of Minas Gerais and São Paulo

Diarrhea in suckling piglets caused by Clostridium perfringens type A was diagnosed in industrial (technified) swine farms of the states of Minas Gerais and São Paulo (Brazil), based on isolation and identification of bacterium by biochemical tests, detection of alpha toxin in animal bioassays, and PCR. This seems to be the first report of clostridial enterotoxaemia in piglets by C. perfringens type A in Brazil and allowed specific procedures to control the disease.

Freqüência de Malassezia pachydermatis em otite externa de cäes / Frequency of Malassezia pachydermatis in canine external otitis

The occurrence of Malassezia pachydermatis in the ears of dogs with otitis externa, comparing both direct exam and growth on culture method was evaluated. Auditory cerumen/exsudate specimens of 50 dogs with otitis externa were analized for M.pachydermatis. Stained by Giemsa, were prepared and microscopically analysed for budding yeasts. Each sample was cultured on modified Sabouraud glucose agar incubated at 28ºC (82.4 F) for detection of M. pachydermatis. The frequency of M. pachydermatis from auditory meatus of otitic dogs was 79 percent (direct microscopy) and 88 percent (growth on modified Sabouraud glucose agar), and no difference between the results provided by both methods used in the fungus detection was observed

An outbreak of acute bovine mastitis caused by Klebsiella pneumoniae in a dairy herd

Neste trabalho está descrito um surto de mamite fatal aguda produzida por Klebsiella pneumoniae em um rebanho leiteiro, localizado no Estado do Rio de Janeiro. Durante quatro meses foram observados 14 mortes de vacas em lactaçäo. Os sinais clínicos observados foram de enterotoxemia aguda e as mortes ocorriam entre 6 e 12 horas após o diagnóstico de mamite por meio do California mastitis test (CMT). Outros microrganismos como Staphylococcus aureus e Streptococcus agalactiae também foram isolados, entretanto ele näo foram associados aos casos agudos e fatais de mamite. O controle inclui medidas de higiene, tratamento de casos clínicos e terapia no período seco, além da vacinaçäo preventiva utilizando-se uma vacina produzida com amostras de K. pneumoniae isoladas do rebanho

Thrombosis of a mitral valve prosthesis resulting from Staphylococcus epidermidis endocarditis.

A 70-year-old man with a Duromedics mitral valve prosthesis had two episodes of infective endocarditis caused by enterococcus (1994 and 1996). Colonoscopy revealed five polyps. Surgical resection was performed and 2 days later the patient had dyspnea and fever. Because of a suspected valve thrombosis, intravenous heparin was given which resulted in hematic effusion in the Douglas' cul-de-sac. Intravenous heparin was withdrawn but the patient continued to have a worsening dyspnea, hyperthermia and hypotension. The patient was transferred to our Institution in cardiogenic shock. Acute thrombosis of the valve was diagnosed by echocardiography, and the patient died before transesophageal Doppler echocardiography was performed. Post mortem examination revealed mitral valve infective thrombosis. In patients with valvular prostheses, endocarditis is an added thromboembolic risk.

Avaliaçäo do teste de imunodifusäo mediante emprego do polissacarídeo "O" no diagnóstica da brucelose bovina / Evaluation of agar gel immunodiffusion test using O-chain polysaccharide for diagnosis of bovine brucellosis

Comparou-se o teste de imunodifusäo dupla em ágar gel (IDGA) pelo emprego do polissacarídeo "O" como antígeno com os testes de soroaglutinaçäo rápida, soroaglutinaçäo lenta, 2-mercaptoetanol, reaçäo de fixaçäo de complemento e antígeno tamponado acidificado no diagnóstico da brucelose, em bovinos infectados, näo infectados e recém-vacinados com a amostra B-19, visando à diferenciaçäo de anticorpos vacinais daqueles associados à infecçäo natural pela Brucella abortus. Nenhum dos testes realizados permitiu diferenciar precisamente anticorpos vacinais daqueles induzidos pela infecçäo brucélica e, nas condiçöes do presente trabalho, a utilizaçäo do IDGA com este objetivo poderia levar à retençäo de animais infectados no rebanho

Isolamento e teste de susceptibilidade a antimicrobianos de bactérias em infecçöes uterinas de éguas / Isolation and antimicrobial susceptibility of bacteria in uterine infections in mares

Foram examinados 206 "swabs" cervicais e uterinos de éguas de várias raças, de diversas regiöes do Estado de Minas Gerais, durante o período de 1986 a 1996. Cerca de 164 "swabs" foram positivos para a presença de microrganismos causadores de endometrites. Streptococcus equi subsp. zooepidemicus (25,7 por cento) e Escherichia coli (15,1 por cento) foram os principais agentes infecciosos isolados. Outros microrganismos presentes foram: Staphylococcus aureus (9,2 por cento), Streptococcus alfa-hemolítico (9,2 por cento), Pseudomonas aeruginosa (3,9 por cento), Staphylococcus coagulase negativo (6,3 por cento), Bacillus spp. (1,9 por cento), Rhodococcus equi (3,4 por cento) e Proteus mirabilis (1,5 por cento). As provas de susceptibilidade aos antimicrobianos revelaram que amicacina e gentamicina (70,2 por cento), ampicilina (59,5 por cento) e cloranfenicol (59,5 por cento) foram os antibióticos de maior açäo in vitro contra os microrganismos isolados

[Benign monoclonal gammopathies: probable antibody specificity of monoclonal immunoglobulins]. / Gammopatie monoclonali benigne: probabile specificità anticorpale delle immunoglobuline monoclonali.

40 patients with benign paraproteinemia have been studied in relation to their age and associated diseases. Significantly high frequency of liver diseases (CALD, cirrhosis, hepatoma, metastases) has been found (12 over 40 people) and increased incidence of idiopathic paraproteinaemia in the old age has been confirmed. 9 patients have been followed for 5 years, so that one could be sure that they had really benign paraproteinaemia: these patients have been then studied from an immunological point of view, in vivo by means of skin tests (PPD, Candida, Trichophyton, DNCB) and in vitro by searching for circulating immune complexes (using a new highly specific immuno-enzymatic method), and compared to controls without paraproteinaemia. Highly positive skin tests have been found only in 7 over 9 patients (even in old subjects) and 6 of them had circulating immune complexes (C.I.C.) in their sera; all the controls were negative both for skin tests and for C.I.C. Immune complexes have been found also in some cases of idiopathic paraproteinaemia, so that they do not seem to be in relation to the associated diseases. The Authors suggest that a genetically determined defect in regulator/suppressor T lymphocyte activity may cause the growth of a benign B cell neoplasm; and that monoclonal immunoglobulins most probably have antibody specificity and are directed against target antigens.