Laboratory Infection of Novel Akhmeta Virus in CAST/EiJ Mice.

Akhmeta virus is a zoonotic Orthopoxvirus first identified in 2013 in the country of Georgia. Subsequent ecological investigations in Georgia have found evidence that this virus is widespread in its geographic distribution within the country and in its host-range, with rodents likely involved in its circulation in the wild. Yet, little is known about the pathogenicity of this virus in rodents. We conducted the first laboratory infection of Akhmeta virus in CAST/EiJ Mus musculus to further characterize this novel virus. We found a dose-dependent effect on mortality and weight loss (p < 0.05). Anti-orthopoxvirus antibodies were detected in the second- and third-highest dose groups (5 × 104 pfu and 3 × 102 pfu) at euthanasia by day 10, and day 14 post-infection, respectively. Anti-orthopoxvirus antibodies were not detected in the highest dose group (3 × 106 pfu), which were euthanized at day 7 post-infection and had high viral load in tissues, suggesting they succumbed to disease prior to mounting an effective immune response. In order of highest burden, viable virus was detected in the nostril, lung, tail, liver and spleen. All individuals tested in the highest dose groups were DNAemic. Akhmeta virus was highly pathogenic in CAST/EiJ Mus musculus, causing 100% mortality when ≥3 × 102 pfu was administered.

Parameters for evaluation of clinical trial in mice infected with Trypanosoma cruzi / Parâmetros para avaliação clínica de camundongos infectados pelo Trypanosoma cruzi em experimentação

The effectiveness of clinical parameters in the evaluation of Trypanosoma cruzi infection was analyzed in male Swiss mice at 8 weeks old Animals were divided into HG (healthy) and IG (1400 trypomastigotes, intraperitoneally, Y strain - Trypanosoma cruzi). Quantitative and qualitative parameters were evaluated in non-consecutive days in the period, from 7th-11th and 15th-18th days of infection. There were significant differences (P<0.05) between both groups in both periods regarding water consumption, abdominal circumference and weight. The second group presented differences in amount of excreta, body temperature, move-up and mortality. There was no difference (P>0.05) between the groups in food consumption, exploration of self-cleaning and skin staining. The fecal feature differed between the groups in the second period. The occurrence of isolation was not practical. Differences were observed in the hair between groups, although the parameter had been interfered by fights between animals. The consumption of water, feed, excreta production, characteristic of the faeces, body temperature, abdominal circumference, move up, weight and mortality parameters are easy to be measured and effective in clinical differentiation of healthy mice infected with T. cruzi, elected in protocols for clinical study with mice, which is the first work to gather information of qualitative and quantitative clinical parameters evaluated in these animals.

Analisou-se a eficiência de parâmetros clínicos na avaliação da infecção pelo Trypanosoma cruzi em camundongos suíços, machos de 8 semanas. Os grupos foram divididos em GS (sadios) e GI (1400 tripomastigotas, intraperitoneal, cepa Y - Trypanosoma cruzi). Avaliaram-se parâmetros quantitativos e qualitativos em dias não consecutivos nos períodos, 7º-11º e 15º-18º dias de infecção. Observaram-se diferenças (P<0.05) significativas entre os grupos, nos dois períodos: consumo de água, circunferência abdominal e peso; apenas no segundo período: quantidade de excretas, temperatura corporal, movimento-levantar e mortalidade. Não houve diferença (P>0.05) entre os grupos: consumo de ração, exploração de auto-limpeza e coloração da pele. As fezes diferiram entre os grupos no segundo período. A ocorrência de isolamento não se mostrou prática. Diferenças no pêlo foram observadas entre os grupos, embora o parâmetro sofra interferência de brigas entre os animais. O consumo de água, ração, produção de excretas, característica das fezes, temperatura corporal, circunferência abdominal, movimento-levantar, peso e mortalidade são parâmetros fáceis de serem medidos e eficientes na diferenciação da clínica de camundongos sadios e infectados pelo T. cruzi, eleitos para protocolos de estudos clínicos com camundongos, sendo este o primeiro trabalho a reunir informações de parâmetros clínicos qualitativos e quantitativos avaliados nesses animais.

Comparison of the sensitivity of in vivo antibody production tests with in vitro PCR-based methods to detect infectious contamination of biological materials.

Bacteria and viruses may be transmitted to laboratory rodents by contaminated biological materials such as transplantable tumours, cell lines, sera or other biological materials. Biological materials are currently being screened using the mouse or rat antibody production (MAP/RAP) test (serological testing). We decided to test and validate an alternative assay using polymerase chain reaction (PCR/realtime PCR) technology to detect viral contamination directly in biological material. The aim of this study therefore is the validation of our new PCR assays and the comparison of PCR and the MAP test. For 8/14 viruses, conventional PCR was more sensitive and more specific than the MAP test in detecting murine viruses. For 12/14 viruses, the realtime PCR was more sensitive than the MAP test. In 2/14 cases, all three detection methods had the same sensitivity. Furthermore, PCR screening clearly conforms to the principles of the 3Rs as a replacement technique because it eliminates the need for using animals to screen for murine viruses in biological material.

Experimental pneumonia virus of mice infection of guineapigs spontaneously infected with Bordetella bronchiseptica.

Guineapigs that were intranasally inoculated with pneumonia virus of mice (PVM) seroconverted to PVM by 11 days post-infection. During the course of study (2-60 days post-infection) no gross or histologic lesions were identified within the lungs that could be attributed to PVM infection. Mild rhinitis and tracheitis were found in most animals and acute purulent bronchopneumonia in two animals, which may have resulted from spontaneous subclinical Bordetella bronchiseptica infection. Viral and bacterial respiratory diseases of the guineapig are briefly reviewed.

Tratamiento de la coccidioidomicosis experimental en ratas Wistar / Experimental treatment of coccidioidomycosis in Wistar rats

Se realizó un estudio comparativo entre Itraconazol (I) y Fluconazol (F) en el tratamiento de la coccidioidomicosis experimental en ratas Wistar. Treinta animales de 250 g de peso fueron inoculados con 200 artroconidias de Coccidioides immitis por vía intercardíaca. Ambas drogas fueron administradas por gastroclisis a razón de 200 mg//kg/día durante 5 semanas a partir del 7 día post-inoculación. El grupo control recibió el solvente de las drogas. Los animales que sobrevivieron 2 semanas después de concluído el tratamiento fueron sacrificados y se le realizaron los siguientes estudios: 1) examen macroscópico (presencia de granulomas) y microscópico (presencia de esporangios) en los pulmones; 2) unidades formadoras de colonias en homogeneizado de pulmones; 3) estudio histopatológico de pulmones con H & E realizando una evaluación semi-cuantitativa contando: a) número de granulomas por campo de 50x; b) número de esporangios con esporangiosporos en cada granuloma y c) número de esporangios vacíos. Los resultados obtenidos mostraron que el itraconazol fue más eficaz que el fluconazol en el tratamiento de este modelo experimental, en la evaluación efectuada a través de varios de los parámetros considerados: 1) se comprobaron menor cantidad de cultivos pulmonares positivos, (30 por ciento vs 100 por ciento); 2) menor número de UFC/g de pulmón (<1x10 vs 1.7x10 ); 3) menos cantidad de granulomas pulmonares en el estudio histopatológico (1/14 campos vs 1/4 campos); 4) menor cantidad de esporangios por granuloma (0 vs 33) y 5) mayor número de esporangios vacíos o alterados. Cabe señalar, sin embargo, que el fluconazol fue capaz de reducir el número de UFC/g pulmón,comparando con el grupo control

Tratamiento de la coccidioidomicosis experimental en ratas Wistar / Experimental treatment of coccidioidomycosis in Wistar rats

Se realizó un estudio comparativo entre Itraconazol (I) y Fluconazol (F) en el tratamiento de la coccidioidomicosis experimental en ratas Wistar. Treinta animales de 250 g de peso fueron inoculados con 200 artroconidias de Coccidioides immitis por vía intercardíaca. Ambas drogas fueron administradas por gastroclisis a razón de 200 mg//kg/día durante 5 semanas a partir del 7 día post-inoculación. El grupo control recibió el solvente de las drogas. Los animales que sobrevivieron 2 semanas después de concluído el tratamiento fueron sacrificados y se le realizaron los siguientes estudios: 1) examen macroscópico (presencia de granulomas) y microscópico (presencia de esporangios) en los pulmones; 2) unidades formadoras de colonias en homogeneizado de pulmones; 3) estudio histopatológico de pulmones con H & E realizando una evaluación semi-cuantitativa contando: a) número de granulomas por campo de 50x; b) número de esporangios con esporangiosporos en cada granuloma y c) número de esporangios vacíos. Los resultados obtenidos mostraron que el itraconazol fue más eficaz que el fluconazol en el tratamiento de este modelo experimental, en la evaluación efectuada a través de varios de los parámetros considerados: 1) se comprobaron menor cantidad de cultivos pulmonares positivos, (30 por ciento vs 100 por ciento); 2) menor número de UFC/g de pulmón (<1x10 vs 1.7x10 ); 3) menos cantidad de granulomas pulmonares en el estudio histopatológico (1/14 campos vs 1/4 campos); 4) menor cantidad de esporangios por granuloma (0 vs 33) y 5) mayor número de esporangios vacíos o alterados. Cabe señalar, sin embargo, que el fluconazol fue capaz de reducir el número de UFC/g pulmón,comparando con el grupo control

An epizootic of Klebsiella aerogenes infection in laboratory rats.

Klebsiella aerogenes was identified as the pathogen in an epizootic of single and multiple abscesses in the cervical, axillary and inguinal regions in laboratory rats. Involvement of the salivary glands in the inflammatory tissue in some cases produced interlobular and interacinar fibrosis resembling "fibroadenosis'. Similar lesions were reproduced with a strain of Klebsiella aerogenes isolated from a diseased rat. Oral carriage of the same strain was detected in the diseased rats, and in normal rats of the same stock. Lymphadenitis appears to have been the primary lesion.