Efficacy of oregano and rosemary essential oils to affect morphology and membrane functions of noncultivable sessile cells of Salmonella Enteritidis 86 in biofilms formed on stainless steel.

AIMS: This study evaluated the efficacy of essential oil from Origanum vulgare L. (oregano; OVEO) and Rosmarinus officinalis L. (rosemary; ROEO) to inactivate sessile cells of Salmonella enterica serovar Enteritidis 86 (SE86) in young and mature biofilms formed on stainless steel. METHODS AND RESULTS: Ultrastructural alterations and damage in different physiological functions caused by OVEO and ROEO in noncultivable sessile cells of SE86 were investigated using scanning electron microscopy and flow cytometry. OVEO (2·5 µl ml-1 ) and ROEO (40 µl ml-1 ) were effective to eradicate young and mature biofilms formed by SE86 sessile cells on stainless steel surfaces; however, the efficacy varied with exposure time. OVEO and ROEO caused alterations in morphology of SE86 sessile cells, inducing the occurrence of bubbles or spots on cell surface. OVEO and ROEO compromised membrane polarization, permeability and efflux activity in noncultivable SE86 sessile cells. These findings show that OVEO and ROEO act by a multitarget mechanism on SE86 membrane functions. CONCLUSIONS: ROEO and OVEO showed efficacy to eradicate SE86 sessile cells in preformed biofilms on stainless steel, displaying a time-dependent effect and multitarget action mode on bacterial cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides for the first time the effects of OVEO and ROEO on morphology and physiological functions of noncultivable sessile cells of S. Enteritidis biofilms preformed on stainless steel surfaces.

Imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas / Immunodetection of Toxoplasma gondii in placenta tissue of naturally infected goats

A imuno-histoquímica (IHQ) é considerada uma ferramenta rápida e precisa para a identificação de protozoários, como Toxoplasma gondii, em tecidos fetais e placentários. Neste estudo foi avaliada a imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas. Foram coletadas e analisadas 80 amostras de placentas de cabras procedentes de único rebanho com sorologia positiva para T. gondii na técnica de ELISA. Na histopatologia, 27/80 amostras apresentaram lesões sugestivas de infecção por protozoários. Após a avaliação histopatológica, procedeu-se à realização da técnica de imuno-histoquímica, obtendo-se 85,2% (23/27) de amostras com marcação positiva. A imunodetecção ocorreu no epitélio de revestimento das vilosidades coriônicas e foi classificada de acordo com o grau de intensidade da imunomarcação. Também foi evidenciada imunomarcação no interior dos vasos sanguíneos fetais em 8,69% (2/23) das amostras. Este estudo demonstrou que a técnica de IHQ se comportou como uma ferramenta valiosa no diagnóstico da infeção por T. gondii em tecido placentário de cabras naturalmente infectadas e complementou, de forma decisiva, o diagnóstico, além de agregar maior valor aos resultados obtidos nas análises histopatológica e sorológica.(AU)

Immunohistochemistry (IHC) is considered to be a rapid and accurate tool for the identification of protozoa such as Toxoplasma gondii in fetal and placental tissues. In this study, we evaluated the immunodetection of Toxoplasma gondii in placental tissue from naturally infected goats. A total of 80 samples of goat placentas from a single herd with positive ELISA serology for T. gondii were collected and analyzed. In the histopathology, 27/80 samples presented lesions suggestive of protozoal infection. After the histopathological evaluation, the immunohistochemistry technique was performed, obtaining 85.2% (23/27) of samples with positive marking. Immunodetection occurred in the lining epithelium of the chorionic villi and was classified according to the degree of intensity of the immunostaining. Immunostaining within the fetal blood vessels was also evidenced in 8.69% (2/23) of the samples. This study demonstrated that the IHQ technique behaved as a valuable tool in the diagnosis of T. gondii infection in placental tissue of naturally infected goats completing the diagnosis in a decisive way and adding greater value to the results obtained in the histopathological and serological analysis.(AU)

Effect of pentoxifylline on the regeneration of rat testicular germ cells after heat shock

High temperature is an important factor for reproduction and can induce testicular degeneration. Pentoxifylline is a methylxanthine phosphodiesterase inhibitor with anti - inflammatory and anti - apoptotic properties. Considering the protective propert ies of pentoxifylline and the harmful effects of heat, the present study aimed to use pentoxifylline to reduce the damage induced by heat to the testis. Adult mal e Wistar rats were exposed to testicular heat shock (43ºC for 15 min) , treated with 50 or 100 mg/ k g of pentoxifylline and evaluated at 3, 7, 15, 30 and 60 days after heat shock. Pentoxifylline treatment did not change testicular weight, histomorphometrical parameters or plasma testosterone concentration. However, pentoxifylline inhibited germ cell apoptosis and reduced the severity of pathological lesions at 30 and 60 days after testicular heat shock. In conclusion, pentoxifylline treatment seem ed to inhibit pro - inflammatory and apoptotic mechanisms triggered by testicular heat shock, improving sper matogenesis regeneration.

Testicular testosterone: estradiol ratio in domestic cats and its relationship to spermatogenesis and epididymal sperm morphology.

The phenomenon of teratozoospermia in felids is not fully understood. In this study, we investigated the testicular androgen:estrogen balance in domestic cats and correlated these data with epididymal sperm morphology and the degree of spermatogenic activity. During spring and summer, testes and blood samples were obtained from 37 mixed-breed domestic cats (12 to 48 mo). The epididymal sperm were harvested and evaluated for sperm counts, motility, and morphology. Distal cytoplasmic droplets were not considered a defect, and samples were considered normozoospermic if they contained more than 60% normal sperm (N = 25) or teratozoospermic if they contained less than 45% normal sperm (N = 12). The testicular and serum concentrations of testosterone (T) and 17ß-estradiol (E2) were determined with an enzyme immunoassay. The gonadosomatic index and epididymal sperm numbers and motility did not differ between groups. The percentage of normal sperm was higher in normozoospermic (74.3 ± 2.0, mean ± SEM) than in teratozoospermic samples (43.1 ± 1.4). The most prevalent sperm defects in the teratozoospermic group were abnormal acrosomes (9.7 ± 2.0) and bent midpieces (12.2 ± 2.0) or tails (24.0 ± 2.7) with cytoplasmic droplets. Histomorphometric data were similar between groups, although there was a lower Leydig cell nuclear volume in teratozoospermic samples. Normozoospermic samples contained a higher percentage of haploid cells and had a higher index of total spermatogenic transformation than teratozoospermic samples. Serum concentrations of T (0.5 ± 0.1 vs. 0.8 ± 0.4 ng/mL) and E2 (9.5 ± 1.2 vs. 11.4 ± 2.3 pg/mL) and testicular T concentrations (471.6 ± 65.3 vs. 313.4 ± 57.6 ng/g) were similar between groups. However, compared with normozoospermic samples, teratozoospermic samples had higher testicular E2 concentrations (8.5 ± 3.6 vs. 5.4 ± 0.5 ng/g) and a lower T:E2 ratio (31.8 ± 4.1 vs. 87.2 ± 11.6). There were significant correlations between testicular E2 values and percentages of normal sperm (r = -0.55) as well as those with primary sperm defects (r = 0.58) or abnormal acrosomes (r = 0.64). The T:E2 ratio was also correlated with meiotic index (r = 0.45) and percentage of normal sperm (r = 0.58). In conclusion, a high testicular E2 concentration and a reduced T:E2 ratio were significantly associated with higher ratios of abnormal sperm types, suggesting that the balance between androgens and estrogens is an important endocrine component in the genesis of teratozoospermia in felids.

Efeito do uso de pentoxifilina no período neonatal sobre a produção espermática em ratos Wistar adultos / Effect of pentoxifylline during neonatal period on spermatic production in adult Wistar rats

Utilizaram-se doses crescentes de pentoxifilina em ratos Wistar neonatos visando aumentar a produção espermática em animais adultos. Trinta e sete animais foram distribuídos de acordo com os tratamentos: não tratados (n=10) e tratados com 1mg/kg (n=10), 5mg/kg (n=9) e 10mg/kg (n=8) de pentoxifilina (IP). Aos 90 dias, os animais foram anestesiados e perfundidos intracardiacamente com solução fixadora. Os testículos foram processados rotineiramente para inclusão em resina plástica à base de glicol metacrilato. Cortes histológicos de 4µm de espessura foram corados em azul de toluidina/borato de sódio a 1 por cento e analisados histometricamente. O número de células de Sertoli por secção transversal diminuiu nos grupos tratados com 5mg/kg e 10mg/kg em relação aos grupos controle e tratado com 1mg/kg. O índice de células de Sertoli aumentou nos animais tratados com 5mg/kg em comparação aos do grupo-controle. A utilização da pentoxifilina não foi capaz de induzir aumento na população das células de Sertoli e produção espermática em ratos adultos.

Increasing doses of pentoxifylline were administrated to newborn Wistar rats in order to augment Sertoli cell number and sperm production in the adult rats. Thirty-seven neonate Wistar rats were distributed in four groups: control (n=10) and treated with 1mg/kg (n=10), 5mg/kg (n=9), and 10mg/kg (n=8) of pentoxifylline. At 90 days, the animals were submitted to anesthesia and intracardiac perfusion. Testes were colleted and routinely processed for inclusion in plastic resin with glycol methacrylate. Histological sections (4µm) were stained in toluidine blue/sodium borate (1 percent) and analyzed. Number of Sertoli cell per transversal section of seminiferous tubule had significant reduction in the groups treated with 5mg/kg and 10mg/kg of pentoxifylline as compared to control and the group that received 1mg/kg (P<0.05). The Sertoli cell index significantly increased in the group treated with 5mg/kg compared to control group. Pentoxifylline did not cause increase in the number of Sertoli cells and daily sperm production in adult male rats.

Sterile nodular dermatitis in dogs.

The types of canine sterile nodular dermatitis discussed in this article have in common the clinical presentation of nodules or plaques; however, they differ in many aspects such as breed predilection, distribution and evolution of cutaneous lesions, systemic involvement, response to therapy, and prognosis. The definitive diagnosis should be based on multiple skin biopsy results. Other ancillary tests may be indicated in cases of systemic involvement. In addition, serum alpha1 antitrypsin can be measured to demonstrate the association between nodular panniculitis and serum alpha1 antitrypsin deficiency. A better understanding of the etiopathogenesis of each of these interesting skin conditions necessitates extensive and systematic diagnostic approaches.

Resolution of superficial necrolytic dermatitis following excision of a glucagon-secreting pancreatic neoplasm in a dog.

An 11-year-old, neutered male standard poodle was diagnosed with superficial necrolytic dermatitis and a glucagon-secreting pancreatic islet neoplasm based on clinical, biochemical, histopathological, immunohistochemical, and hormonal findings. Hyperglucagonemia, hyperinsulinemia, and hypoaminoacidemia were observed on preoperative laboratory analysis. Abnormal laboratory values returned to normal, and complete resolution of skin lesions occurred after tumor excision. The dog has remained clinically normal for six months following surgery.

Melanoma in situ on facial skin damaged by sunlight.

Solar lentigines ("liver spots") commonly occur on sun-damaged skin in the elderly population. Although they are benign, it is difficult in practice to distinguish solar letigines and junctional nevi from melanoma in situ. We report a patient who developed melanoma on the same site as a previously diagnosed solar lentigo. We also review the histopathological criteria for differentiating these benign lesions from potentially malignant ones.

Cutaneous plasmacytic infiltrates.

This article reviews a group of skin disorders that have in common the presence of a cellular infiltrate with plasma cells upon histopathologic examination. These include inflammatory and neoplastic diseases and a group of miscellaneous disorders of varied etiology. The correlation between the presence of plasma cells in the cellular infiltrate and the diagnosis, prognosis, and pathogenesis of the disorders is emphasized.