Intrapuparial development of Sarcophaga (Liopygia) ruficornis (Diptera: Sarcophagidae), a species of medical-veterinary and forensic importance, under laboratory conditions.

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) is a species of medical-veterinary and forensic importance, as its immatures cause myiasis in humans and animals and colonize carcasses and cadavers. Therefore, investigations into the biology and morphology of this species, with a particular focus on pupae that constitute ≥50% of the developmental period for the immatures, are pertinent. Although there are biological and morphological studies of pupae, the intrapuparial development at different temperatures has not yet been analysed. Therefore, the present study aimed to describe how temperature (22, 27 and 32 ± 1°C) affects the development and morphology of S. (L.) ruficornis pupae at 60 ± 10% relative humidity and a 12-h photoperiod. Ten pupae were collected, euthanized and fixed every 4 h from pupariation until 24 h and every 8 h until the emergence of the first adult. Emergence occurred at 440, 272 and 232 h at 22, 27 and 32°C, with 590, 380 and 330 pupae, respectively. The highest mortality occurred at 32°C. Eight periods were defined, and sex was determined in pharate adult stage; in addition, 40 key morphological characteristics to estimate pupal age were presented. These findings can serve as a basis for studies on the biology and morphology of the pupa of S. (L.) ruficornis, particularly in bionomics, control and forensics, helping researchers and experts.

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) é uma espécie de importância médico­veterinária e forense, pois seus imaturos causam miíase em humanos e animais e colonizam carcaças e cadáveres. Portanto, são pertinentes as investigações sobre a biologia e morfologia dessa espécie, com foco especial nas pupas que constituem ≥50% do período de desenvolvimento dos imaturos. Embora existam estudos biológicos e morfológicos das pupas, o desenvolvimento intrapuparial em diferentes temperaturas ainda não foi analisado. Logo, o presente estudo teve como objetivo descrever como a temperatura (22, 27 e 32 ± 1°C) afeta o desenvolvimento e a morfologia das pupas de S. (L.) ruficornis a 60 ± 10% de umidade relativa e fotoperíodo de 12 horas. Dez pupas foram coletadas, eutanasiadas e fixadas a cada quatro horas desde a pupariação até 24 horas e a cada oito horas até a emergência do primeiro adulto. A emergência ocorreu em 440, 272 e 232 horas a 22, 27 e 32°C, com 590, 380 e 330 pupas, respectivamente. A maior mortalidade ocorreu a 32°C. Foram definidos oito períodos e o sexo foi determinado na fase de adulto farato; além disso, foram apresentadas 40 características morfológicas importantes para estimar a idade das pupas. Essas descobertas podem servir de base para estudos sobre a biologia e a morfologia da pupa de S. (L.) ruficornis, especialmente em bionomia, controle e ciência forense, ajudando pesquisadores e especialistas.

The Power of Discourse: Associations between Trainers' Speech and the Responses of Socialized Wolves and Dogs to Training.

In a previous study, we found that Positive Reinforcement Training reduced cortisol of wolves and dogs; however, this effect varied across trainer-animal dyads. Here we investigate whether and how the trainers' use of speech may contribute to this effect. Dogs' great interest in high-pitched, intense speech (also known as Dog Directed Speech) has already been reported, but whether and how wolves respond similarly/differently to voice characteristics has never been studied before. We analyzed 270 training sessions, conducted by five trainers, with nine mixed-breed dogs and nine wolves, all human-socialized. Through Generalized Linear Mixed Models, we analyzed the effects of (a) three speech categories (nice, neutral, reprehensive) and laugh; and (b) acoustic characteristics of trainers' voices on animals' responses (correct responses, latency, orientation, time at less than 1 m, non-training behaviors, tail position/movements, cortisol variation). In both subspecies, tail wagging occurred more often in sessions with longer durations of nice speech, and less often in sessions with reprehensive speech. For dogs, the duration of reprehensive speech within a session was also negatively related to correct responses. For wolves, retreat time was associated with more reprehensive speech, whereas duration of nice speech was positively associated with time spent within one meter from the trainer. In addition, most dog behavioral responses were associated with higher average intonations within sessions, while wolf responses were correlated with lower intonations within sessions. We did not find any effects of the variables considered on cortisol variation. Our study highlights the relevance of voice tone and speech in a training context on animals' performances and emotional reactions.

Acoustic Monitoring of Black-Tufted Marmosets in a Tropical Forest Disturbed by Mining Noise.

All habitats have noise, but anthropogenic sounds often differ from natural sounds in terms of frequency, duration and intensity, and therefore may disrupt animal vocal communication. This study aimed to investigate whether vocalizations emitted by black-tufted marmosets (Callithrix penicillata) were affected by the noise produced by mining activity. Through passive acoustic monitoring, we compared the noise levels and acoustic parameters of the contact calls of marmosets living in two study areas (with two sampling points within each area)-one near and one far from an opencast mine in Brazil. The near area had higher anthropogenic background noise levels and the marmosets showed greater calling activity compared to the far area. Calls in the near area had significantly lower minimum, maximum and peak frequencies and higher average power density and bandwidth than those in the far area. Our results indicate that the mining noise affected marmoset vocal communication and may be causing the animals to adjust their acoustic communication patterns to increase the efficiency of signal propagation. Given that vocalizations are an important part of social interactions in this species, concerns arise about the potential negative impact of mining noise on marmosets exposed to this human activity.

Development and Intrapuparial Characterization of Hydrotaea aenescens (Diptera: Muscidae) Raised at Different Temperatures Under Laboratory Conditions.

Species belonging to the Muscidae family are important in the medical-sanitary and forensic fields due to their biology. The intrapuparial stage of Hydrotaea aenescens (Wiedemann, 1830), which represents about 50% of the immature development cycle in muscoid Diptera, has not yet been studied in detail. This study identifies and describes the main temporal morphological changes that occur in the pupal body of H. aenescens during its development at 22, 27, and 32 ± 1°C, RH 60 ± 10% with a 12-hr photoperiod. Ten pupae were collected, sacrificed, and fixed every 3 hr after pupariation up to 24 hr, and every 6 hr thereafter until the first emergence. Emergence took place at 282, 174, and 126 hr at 22, 27, and 32°C, with 520, 340, and 260 pupae, respectively. External morphology was described and recorded at eight development stages: pupariation, pre-pupae, early and late cryptocephalic pupae, phanerocephalic pupae, pupae-adult apolysis, pharate adults, and imago, employing 18 key morphological characters for pupae age. These morphological pupae characters, comprise, therefore an alternative and/or additional method to aid in determining the minimum Postmortem Interval (minPMI).

Scale-free distribution of silences.

Soundscape studies help us understand ecological processes, biodiversity distribution, anthropic influences, and even urban quality, across a wide variety of places and time periods. In this work, instead of looking for differences, we ask if there are common characteristics shared by all soundscapes. Based on our results, we propose a universal distribution of quiet-time (background noise) and sound-time (acoustic energy bursts) in audio recordings. We analyzed one continuous hour during daylight and one at night, from ten randomly selected days in each environment: urban, dry forest, savanna, rupestrian field, Atlantic forest, marine, and freshwater. We found that the histograms of the quiet-time followed a power law for all scenarios analyzed, they present fractal events or scale-free distributions. This distribution covers up to four orders of magnitude, with an exponent of 1.6≤α≤2.0 for all soundscapes. By contrast, the sound-time distribution in all environments followed a log-normal or timescale dependence, with a typical time for the duration of sounds (0.06-0.12 s). Such time duration limitation can be related to the physiology of sound emission in animals.

Temperature and exudativory as drivers of the marmoset (Callithrix spp.) daily activity period.

Primates are affected by fluctuations in ambient temperatures, mostly through thermoregulatory costs and changes in the availability of food. In the present study, we investigate whether the ambient temperature and proxies of food availability affect the activity period of marmosets (Callithrix spp.). We predicted that: (i) at colder sites, marmosets would spend more time at sleeping sites; (ii) midday resting bouts would be longer at hotter sites; (iii) the onset/cessation of activity and resting behavior at midday would be more closely related to temperature than food availability, and (iv) highly exudativorous groups would have higher total levels of resting. We compiled data on the onset and cessation of activity and the time spent resting at midday from seven marmoset studies from sites with a wide range of temperatures. We used generalized linear mixed models to verify the relationship between the dependent variables (lag between dawn and the onset of activities, lag between cessation of activities and dusk, and proportion of resting during midday) and the minimum and maximum temperatures at the respective study sites, together with proxies of food availability (exudativory rates, the amount of habitat available per individual, and net primary productivity) using each sample month as a sampling unit and the identity of the study as a categorical random factor. At colder sites and during colder months, the marmosets left sleeping trees later in the morning and ceased their activities earlier, while at hotter sites and during hotter months, they spent more time resting during midday. More exudativorous groups become active later in the morning, but also ceased their activities later. The abundance of food did not affect the timing of activities. We provide evidence that both low and high temperatures affect marmosets' activities, and that their activity period appears to be more influenced by the thermal environment than food availability.

Epidemiology of influenza B infection in the state of Rio Grande do Sul, Brazil, from 2003 to 2019.

Influenza B virus (IBV) causes respiratory tract infections with mild, moderate, or life-threatening symptoms. This study describes the epidemiology of IBV infection in Rio Grande do Sul (RS), Brazil, over 17 years. Nasopharyngeal samples were collected from outpatients presenting acute respiratory illness (ARI) between 2003 and 2019, and from inpatients with severe acute respiratory infection (SARI) from 2009 to 2019. IBV was detected by immunofluorescence assay or quantitative real-time polymerase chain reaction; demographic and clinical data were analyzed. In total, 48,656 cases of respiratory infection were analyzed, of which 20.45% were ARI, and 79.46% were SARI. Respiratory viruses accounted for 22.59% and 37.47% of the cases of ARI and SARI, respectively. Considering respiratory viral infections, 17.10% of ARI and 3.06% of SARI were associated with IBV. IBV circulated year-round in RS, with an increase in autumn and winter, peaking in July (p = .005). IBV infection showed an association with age, and most outpatients positive for IBV were between 10 and 49 years old, whereas IBV infection in SARI affected mainly individuals ≤ 1 year or ≥ 60 years old. No significant association was found between sex and IBV infection. Coryza, sore throat, and myalgia were associated with ARI (p < .001). Moreover, 3.18% of the deaths associated with respiratory virus infection were positive for IBV; notably, cardiopathy (p < .001), metabolic disease (p < .001), and smoking (p = .003) were associated to fatality in IBV infection. IBV is an important cause of severe respiratory infections, and the fatality risk is high in individuals with cardiopathy and metabolic diseases.

Estudo exploratório sobre a utilização da Colostroterapia em Unidade Neonatal de uma maternidade brasileira / Exploratory study on the use of Colostrum therapy in a brazilian maternity hospital's Neonatal Unit

Estudo exploratório para verificar a utilização da colostroterapia em recém-nascidos de muito baixo peso, em unidade neonatal de maternidade de médio porte, por meio da avaliação de prontuários. Dos 108 prontuários avaliados, 56,5% dos neonatos eram do sexo feminino, com média de peso e idade gestacional ao nascimento de 1091,9 g (DP=263,7g) e 29 semanas/2dias (DP=2semanas/6dias), respectivamente. A colostroterapia foi utilizada em 25,9% dos pacientes, sendo que em 96,4% dos casos a administração foi por via oral. O uso de terapia colostral foi associado significativamente a variáveis neonatais como idade gestacional, peso ao nascer e óbito neonatal (p = 0,001, p < 0,001, e p < 0,001). Não havendo protocolo estabelecido para guiar a prescrição da colostroterapia, esta ocorreu principalmente para prematuros de menor peso, mais imaturos ou mais doentes, com maior risco de morbidade e mortalidade.

Exploratory study of colostrum therapy use in very low birth weight newborns in a neonatal unit by the analysis of medical records. The 108 patients under study were predominantly female (56,5%), had a mean weight and gestational age at birth of 1091.9 g (SD=263.7g) and 29 weeks/2 days (SD=2 weeks/6 days), respectively. Colostrum therapy was used for 29.5% of the patients mostly by oral administration (96.4%). The use of colostrum therapy was significantly associated with low gestational age and weight at birth and with neonatal death (p = 0.001, p < 0.001 and p < 0.001). As there was no established protocol to guide colostrum therapy prescription in the neonatal unit, the therapy was apparently chosen to be used for more immature or sicker babies, with a greater morbidity or mortality risk.

Effects of mining truck traffic on cricket calling activity.

Anthropogenic noise is a global pollutant and several studies have identified its impact on wildlife. This research shows how the noise produced by mining affects crickets' acoustic communication. Two passive acoustic monitoring devices (SMII) were installed in a forest fragment located at 500 m from the Brucutu Mine in Brazil. Another two SMII were installed distant 2500 from the mine. The equipment was configured to record from 17:00 to 05:00 h during seven days in April 2013. The authors analyzed the spectral characteristics of acoustic activity of three species of crickets (Anaxipha sp., Gryllus sp., and a Podoscirtinae species) before, during, and after the passing of mine trucks. For comparison the authors analyzed the acoustic characteristics for Anaxipha sp. and Gryllus sp. found in the distant site. Results showed a calling interruption for all the species during truck transit. Gryllus sp. emitted calls with higher maximum frequencies, average power, and larger bandwidth in the site close to the mine. Podoscirtinae species emitted calls with lower minimum frequencies, higher average power, and large bandwidth in the close site. The authors show that insect acoustic behavior varies between areas with different levels of noise. The disruption of this behavior may have negative consequences for their reproductive success.

Humoral responses in cattle to commercial vaccines containing Clostridium perfringens epsilon toxoid and C. botulinum types C and D toxoids last less than a-year.

The aim of this study was to evaluate the titers of neutralizing antibodies in cattle inoculated with multivalent commercial clostridial vaccines containing C. botulinum type C (BoNTC), C. botulinum type D (BoNTD), and C. perfringens epsilon (ETX) toxoids for a period of one year. Cattle (Bos taurus), aged 4-6 months and not previously immunized, were vaccinated under four different protocols at days 0 and 30 and followed over one year. Individual serum titration was performed by a serum neutralization test in mice or in MDCK cells. The number of animals with detectable neutralizing antibodies ranged from 40.6% to 78.1%, but only 12.5% of animals showed neutralizing antibodies against all tested antigens. Neutralizing antibodies were found only until 60 days for ETX, 120 days for BoNTC, and 180 days for BoNTD. The absence of detectable neutralizing antibodies against the three antigens before 360 days, suggests that cattle remained unprotected for a long period before the recommended booster vaccination.

Comparison of humoral neutralizing antibody response in rabbits, guinea pigs, and cattle vaccinated with epsilon and beta toxoids from Clostridium perfringens and C. botulinum types C and D toxoids.

Rabbits and guinea pigs are used in the official control and validation of clostridial vaccines, but it is unknown whether the antitoxin titers obtained in these animals corroborate with the humoral response in bovine. The objective of the study was to compare the humoral antibody response of guinea pig and rabbits to those obtained in cattle vaccinated with a commercial vaccine containing Clostridium perfringens epsilon and beta, and Clostridium botulinum types C and D toxoids. This study revealed the same level of humoral response in rabbits and cattle for all four toxoids tested, including C. botulinum types C and D toxoids. In contrast, the titers of neutralizing antibodies against C. botulinum type C toxin in guinea pigs differed from those obtained in cattle. Thus, the present work suggests that the potency test for C. botulinum types C in rabbits agrees more with the humoral response in cattle than the potency test in guinea pigs, thereby making it possible to use only rabbits as models in the official control and validations of clostridial vaccines.

Behavioural and ecological aspects of black tufted-ear marmosets, Callithrix penicillata (Geoffroy, 1812) (Primates: Callitrichidae) in a semi-urban environment

O mico-estrela (Callithrix penicillata) é uma pequena espécie de primata neotropical que vive em vários tipos de ambientes como Cerrado e Mata Atlântica, podendo também ocupar áreas urbanas. Pesquisas já mostraram que a disponibilidade de recurso alimentar afeta a área de vida, o tamanho de grupo e o comportamento da espécie na natureza. Os objetivos deste estudo foram investigar o comportamento do mico-estrela em um ambiente urbanizado e a influência de recurso alimentar (estimado) na área de vida e na densidade populacional. O estudo foi realizado por meio de observações diretas (84 horas) com quatro grupos de mico-estrela vivendo nas dependências do zoológico de Belo Horizonte, Minas Gerais, Brasil. Os resultados mostraram que apesar de viverem em ambiente urbano, estes micos tem áreas de vida (2,85ha), tamanhos dos grupos (média=5,7 indivíduos) e densidades populacionais (2,19 ind/ha) parecidos com os valores obtidos para a espécie nativa. Os animais estudados, passaram a maior parte do dia se deslocando (22,45%), descansando (16,58%), inativos alertas (14,95%) e alimentando-se (6,74%). Estes resultados mostram que o padrão de atividade e o repertório comportamental exibidos pelos animais deste estudo são similares aos apresentados na natureza e portanto, evidenciam a capacidade de adaptação da espécie em ambientes antrópicos.(AU)

The black tufted-ear marmoset (Callithrix penicillata) is a small Neotropical primate, which lives in various types of environments from Cerrado (Brazilian savannah), Atlantic forest to urban areas. Research shows that food availability influences home range size, group size, population density and the behaviour of marmoset groups in the wild. The objective of this study was to investigate the behaviour of C. penicillata in an urbanized habitat, and the influence of (estimated) food availability on home range size, group size and population density. This study was performed through direct behavioural observation (84 hours) on four wild groups of marmosets living within the grounds of Belo Horizonte Zoological garden, Minas Gerais, Brazil. The results show that, despite living in an urban habitat, these marmosets presented home range size (2.85ha), group size (mean=5.7 ind.) and population density (2.19 ind/ha) values in the range found in wild-living conspecifics. The animals studied spent a considerable percentage of their time travelling (22.45%); resting (16.58%), inactive alert (14.95%) and feeding (6.74%). These results showed that behavioural activity repertoires were also found to be similar to wild-living conspecifics and therefore demonstrate the species adaptation capacity in urban environment.(AU)

Behavioural and ecological aspects of black tufted-ear marmosets, callithrix penicillata (geoffroy, 1812) (primates: callitrichidae) in a semi-urban environment

The black tufted-ear marmoset (Callithrix penicillata) is a small Neotropical primate, which lives in various types of environments from Cerrado (Brazilian savannah), Atlantic forest to urban areas. Research shows that food availability influences home range size, group size, population density and the behaviour of marmoset groups in the wild. The objective of this study was to investigate the behaviour of C. penicillata in an urbanized habitat, and the influence of (estimated) food availability on home range size, group size and population density. This study was performed through direct behavioural observation (84 hours) on four wild groups of marmosets living within the grounds of Belo Horizonte Zoological garden, Minas Gerais, Brazil. The results show that, despite living in an urban habitat, these marmosets presented home range size (2.85ha), group size (mean=5.7 ind.) and population density (2.19 ind/ha) values in the range found in wild-living conspecifics. The animals studied spent a considerable percentage of their time travelling (22.45%); resting (16.58%), inactive alert (14.95%) and feeding (6.74%). These results showed that behavioural activity repertoires were also found to be similar to wild-living conspecifics and therefore demonstrate the species adaptation capacity in urban environment.

O mico-estrela (Callithrix penicillata) é uma pequena espécie de primata neotropical que vive em vários tipos de ambientes como Cerrado e Mata Atlântica, podendo também ocupar áreas urbanas. Pesquisas já mostraram que a disponibilidade de recurso alimentar afeta a área de vida, o tamanho de grupo e o comportamento da espécie na natureza. Os objetivos deste estudo foram investigar o comportamento do mico-estrela em um ambiente urbanizado e a influência de recurso alimentar (estimado) na área de vida e na densidade populacional. O estudo foi realizado por meio de observações diretas (84 horas) com quatro grupos de mico-estrela vivendo nas dependências do zoológico de Belo Horizonte, Minas Gerais, Brasil. Os resultados mostraram que apesar de viverem em ambiente urbano, estes micos tem áreas de vida (2,85ha), tamanhos dos grupos (média=5,7 indivíduos) e densidades populacionais (2,19 ind/ha) parecidos com os valores obtidos para a espécie nativa. Os animais estudados, passaram a maior parte do dia se deslocando (22,45%), descansando (16,58%), inativos alertas (14,95%) e alimentando-se (6,74%). Estes resultados mostram que o padrão de atividade e o repertório comportamental exibidos pelos animais deste estudo são similares aos apresentados na natureza e portanto, evidenciam a capacidade de adaptação da espécie em ambientes antrópicos.

Atividade citotóxica do extrato de Vitis labrusca em culturas de Trichomonas vaginalis / Cytotoxic activity of Vitis labrusca extract in cultures of Trichomonas vaginalis

Objetivos: Avaliar a atividade anti-Trichomonas vaginalis do extrato de folhas de videira (Vitis labrusca) provenientes de cultivo convencional e orgânico.Métodos: Extratos de folhas secas de videira, orgânico e convencional, foram elaborados com extrator Sohxlet. A composição fenólica total dos extratos foi avaliada por Folin-Cicoalteau e Cromatografia Líquida de Alta Eficiência. A atividade antiparasitária dos extratos foi realizada em cultura de T. vaginalis. Os testes foram realizados em três concentrações finais: 5, 10 e 20 mg/mL. Os trofozoítos foram contados em hemocitômetro e sua densidade foi ajustada em meio de cultura Trypticase-Yeast Extract-Maltose (TYM), de forma a se obter a concentração final de 1×105 trofozoítos/mL. Após incubação com os extratos por 24 horas a 37°C, foi feita a contagem dos trofozoítos com motilidade em hemocitômetro. Os controles do veículo foram realizados, adicionando-se água em substituição aos extratos.Resultados: O extrato de folhas convencionais é rico em catequina e os orgânicos, em resveratrol. Os extratos de V. labrusca não apresentaram atividade anti-T. vaginalis nas concentrações de 5 e 10 mg/mL. Entretanto na concentração de 20 mg/L, o extrato de folhas secas convencionais reduziu em 100% e o extrato orgânico em 48% a sobrevivência dos trofozoítos. Os polifenóis catequina, quercetina e rutina estavam em maior concentração no extrato com maior eficácia, os quais podem ser os responsáveis pela ação anti-T. vaginalis do extrato convencional.Conclusões: O extrato de folha de videira pode ser uma alternativa no combate a T. vaginalis. Estudos pré-clínicos e clínicos são necessários para comprovar eficácia e segurança desta intervenção...

AIMS: To evaluate the anti-Trichomonas vaginalis activity of the extract of organic and conventional leaves of Vitis labrusca.METHODS: Organic and conventional grapevine leaf extracts were prepared with extractor Sohxlet. The total phenolic extracts were evaluated by Folin-Cicoalteau and High-Performace Liquid Chromatography. The antiparasitic activity of the extracts was performed on growth of T. vaginalis. Tests were conducted at three final concentrations of 5, 10 and 20 mg/mL. The trophozoites were counted by hemocytometer and their density was adjusted in Trypticase-Yeast Extract-Maltose (TYM) growth environment in order to obtain a final concentration of 1x105 trophozoites/ml. After incubated with the extracts for 24 hours at 37 �C, the count of motile trophozoites was taken by hemocytometer. The vehicle controls were performed by substituting the extract by water.RESULTS: The conventional leaf extract is rich in catechin and the organic in resveratrol. The extracts of V. labrusca showed no anti-T.vaginalis activity at concentrations of 5 and 10 mg/mL. However in the concentration of 20 mg/L conventional grapevine leaf extracts reduced by 100% and the organic extract by 48% the survival of trophozoites. Catechin polyphenols, quercetin and rutin were in higher concentration in the extract with greater efficacy, which may be the responsible for the anti-T. vaginalis action of the conventional extract.CONCLUSIONS: The vine leaf extract may be an alternative to combat T. vaginalis. Pre-clinical and clinical studies are needed to confirm efficacy and safety of this intervention...

Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil.

The objective of this study was to detect C. difficile A/B toxins and to isolate strains of C. perfringens and C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficile A/B toxins were detected by ELISA, and C. perfringens and C. difficile were identified by multiplex PCR. C. difficile A/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A(+)B(+) and two were A(-)B(-). All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtB was found in one strain, which was A(+)B(+) and was derived from a non-diarrheic dog. C. perfringens strains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringens type A and there was an association between the detection of the cpe gene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2 gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficile and C. perfringens in dogs and to better our understanding of C. difficile as a zoonotic agent. This is the first study to report the binary toxin gene in C. difficile strains isolated from dogs in Brazil.

Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil

The objective of this study was to detect C. difficileA/B toxins and to isolate strains of C. perfringensand C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficileA/B toxins were detected by ELISA, and C. perfringensand C. difficilewere identified by multiplex PCR. C. difficileA/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A+B+and two were A-B-. All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtBwas found in one strain, which was A+B+and was derived from a non-diarrheic dog. C. perfringensstrains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringenstype A and there was an association between the detection of the cpegene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficileand C. perfringensin dogs and to better our understanding of C. difficileas a zoonotic agent. This is the first study to report the binary toxin gene in C. difficilestrains isolated from dogs in Brazil.

Noisy human neighbours affect where urban monkeys live.

Urban areas and many natural habitats are being dominated by a new selection pressure: anthropogenic noise. The ongoing expansion of urban areas, roads and airports throughout the world makes the noise almost omnipresent. Urbanization and the increase of noise levels form a major threat to living conditions in and around cities. Insight into the behavioural strategies of urban survivors may explain the sensitivity of other species to urban selection pressures. Here, we show that urban black-tufted marmosets (Callithrix penicillata) living in noisy urban areas may select their home-range based primarily on ambient noise level. We have tested the hypothesis that the noise from vehicular traffic and visitors in an urban park in Brazil influences the use of home-range (space) by urban marmosets. Marmosets even avoided noisy areas with high food availability. In addition, they systematically preferred the quieter areas even with dynamic changes in the acoustic landscape of the park between weekdays and Sundays (no observations were made on Saturdays). These data provide evidence that the use of home-range by wild animals can be affected by a potential aversive stimulus such as noise pollution.

Improving efficacy of interleukin-12-transfected dendritic cells injected into murine colon cancer with anti-CD137 monoclonal antibodies and alloantigens.

Intralesional administration of cultured dendritic cells (DCs) engineered to produce IL-12 by in vitro infection with recombinant adenovirus frequently displays eradicating efficacy against established subcutaneous tumors derived from the CT26 murine colon carcinoma cell line. The elicited response is mainly mediated by cytolytic T lymphocytes. In order to search for strategies that would enhance the efficacy of the therapeutic procedure against less immunogenic tumors, we moved onto malignancies derived from the inoculation of MC38 colon cancer cells that are less prone to undergo complete regression upon a single intratumoral injection of IL-12-secreting DCs. In this model, we found that repeated injections of such DCs, as opposed to a single injection, achieved better efficacy against both the injected and a distantly implanted tumor; that the use of semiallogeneic DCs that are mismatched in one MHC haplotype with the tumor host showed slightly better efficacy; and that the combination of this treatment with systemic injections of immunostimulatory anti-CD137 (4-1BB) monoclonal antibody achieved potent combined effects that correlated with the antitumor immune response measured in IFN-gamma ELISPOT assays. The elicited systemic immune response eradicates concomitant untreated lesions in most cases. Curative efficacy was also found against some tumors established for 2 weeks when these strategies were used in combination. These are preclinical pieces of evidence to be considered in order to enhance the therapeutic benefit of a strategy that is currently being tested in clinical trials. Supplementary Material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.