Isolation of Mycoplasma moatsii from the intestine of wild Norway rats (Rattus norvegicus)

The intestinal tracts of twenty inbred SPF rats (LEW, BN, WKY, DA) and six wild Norway rats (Rattus norvegicus Berkenhout 1769) were investigated for mycoplasmas. Cultivation was in three different media. Mycoplasmas were not isolated from the intestine of the inbred SPF rats but were found in the epithelia and contents of caecum, colon and jejunum as well as in fecal samples of all of the wild Norway rats investigated. The mycoplasmas isolated all belonged to the same species, and were identified as Mycoplasma moatsii, originally isolated from grivit monkeys (Cercopithecus aethiops) and thought to be specific for this host.     

Capillaria hepatica infection in wild brown rats (Rattus norvegicus) from the urban area of Milan, Italy

Forty-seven wild brown rats (Rattus norvegicus) collected from the urban area of Milan (Italy) were screened for Capillaria hepatica liver infection. The liver of each rat was grossly and histologically examined for the presence of C. hepatica adults, eggs and typical C. hepatica induced lesions. In 17 rats (36%) liver lesions consistent with C. hepatica infection were detected. Grossly, white-yellow nodules of 1-5 mm in diameter were present, either scattered on the liver surface or localized in a single lobe. Histologically, granulomatous liver lesions associated with eggs and/or worms were observed. The degree of gross liver involvement was moderate in most of the positive cases (71%). About 30 cases of C. hepatica infection in humans have been documented world-wide, most of which are reported in children from 1 to 5 years of age. Our results suggest that the potential transmission of C. hepatica to children in the study area should be considered an important health issue.     

First detection of Hepatitis E virus (Orthohepevirus C) in wild brown rats (Rattus norvegicus) from Great Britain

In the United Kingdom, there has been an increase in the number of hepatitis E virus (HEV) infections in people annually since 2010. Most of these are thought to be indigenously acquired Orthohepevirus A genotype 3 (HEV G3), which has been linked to pork production and consumption. However, the dominant subgroup circulating in British pigs differs from that which is found in people; therefore, an alternative, potentially zoonotic, source is suspected as a possible cause of these infections. Rodents, brown rats (Rattus norvegicus) in particular, have been shown to carry HEV, both the swine HEV G3 genotype and Orthohepevirus C, genotype C1 (rat HEV). To investigate the prevalence of HEV in British rodents, liver tissue was taken from 307 rodents collected from pig farms (n = 12) and other locations (n = 10). The RNA from these samples was extracted and tested using a pan‐HEV nested RT‐PCR. Limited histopathology was also performed. In this study, 8/61 (13%, 95% CI, 5–21) of brown rat livers were positive for HEV RNA. Sequencing of amplicons demonstrated all infections to be rat HEV with 87%–92% nucleotide identity to other rat HEV sequences circulating within Europe and China (224 nt ORF‐1). Lesions and necrosis were observed histologically in 2/3 samples examined. No rat HEV RNA was detected in any other species, and no HEV G3 RNA was detected in any rodent in this study. This is the first reported detection of rat HEV in Great Britain. A human case of rat HEV infection has recently been reported in Asia, suggesting that rat HEV could pose a risk to public health.     

Finding of Neospora caninum in the wild brown rat (Rattus norvegicus)

Nine rats (16.4%) out of 55 (Rattus norvegicus) from cattle farms were seropositive to Neospora caninum. Two of the seropositive rats were also PCR positive but all were negative by immunohistochemistry and PAS staining. The brains of all the captured rats were homogenized and initially inoculated intraperitoneally into nude mice or into SPF ICR mice, which had been immunosuppressed with prednisolone. One mouse that was inoculated with brain material from a seropositive rat became infected with N. caninum, as demonstrated by the presence of a tissue cyst in the brain and confirmed by immunohistochemistry and PCR. This is the first finding of N. caninum in naturally infected farm rats. The findings show that natural N. caninum infection occurs in wild brown rats and thus rats may serve as a reservoir for the protozoan on the cattle farm.     

Efficacy of anticoagulant-free alternative bait products against house mice (Mus musculus) and brown rats (Rattus norvegicus)

The attractiveness and efficacy of four anonymous anticoagulant-free alternative rodenticides (active ingredients cellulose or plaster) were tested on wild strain groups of house mice (Mus musculus Linnaeus) and brown rats (Rattus norvegicus Berkenhout) in laboratory choice and no-choice experiments. In no-choice tests, the rodenticide product was offered ad libitum to the animals for 21 days (mice) and 10 days (rats), respectively. In choice tests, the rodenticide product and non-poisoned food were offered for 28 days (mice) and 14 days (rats). Two products with cellulose (C1 and C2) as the active ingredient were tested against both mice and rats. In two no-choice tests with mice, all animals died within 14 days (C1) and 21 days (C2); in a third no-choice test, 11% (2/18) of mice survived (C1: 21 days). In all three tests, numerous incidents of cannibalism were observed. The attractiveness of cellulose baits was tested for one product (C1) in three trials against normal food pellets. In all trials, bait consumption was low and all mice survived (15–34 days). Two no-choice trials with cellulose baits (C1) were conducted on groups of Rattus norvegicus; 11 of 12 rats survived the bait feeding period and no cannibalism or sign of aggression between conspecifics was observed. Two plaster-based (CaSO₄·0.5H₂O) products (P1 and P2) were tested against both mice and rats; in no-choice tests, only one mouse (n= 20) and no rats (n= 8) were killed, although the baits proved to be attractive for rats and mice in choice tests. The results of these experiments suggest that neither cellulose-based nor plaster-based rodenticides are suitable agents for the control of Mus musculus and Rattus norvegicus.     

Transmission and shedding patterns of Salmonella in naturally infected captive wild roof rats (Rattus rattus) from a Salmonella-contaminated layer farm

Rodents play a major role in the transmission and maintenance of Salmonella contamination cycles in poultry facilities. However, very limited field data are available regarding the transmission routes, infection cycle, and shedding patterns of Salmonella by naturally infected wild rodents from commercial layer farms. In this study, a total of 128 resident wild roof rats (Rattus ratus) were captured from a Salmonella-contaminated layer facility. All roof rats were divided into 51 laboratory cages, and weekly monitoring of Salmonella fecal shedding patterns was conducted for 53 wk. Seven roof rats from cages that were observed to frequently shed Salmonella were isolated in individual cages, and daily Salmonella monitoring was performed for 35 days. At the end of monitoring, each roof rat was euthanatized, and isolation of Salmonella from different organs was performed. Results of weekly monitoring of Salmonella showed that 21 of 51 cages (41.2%) were positive for Salmonella Infantis, while two cages (3.92%) were positive for Salmonella Enteritidis. Moreover, 11 cages were positive for Salmonella for at least two sampling weeks. Isolation of Salmonella from fecal droppings was mainly observed during the first 12 wk of captivity. The longest interval between two Salmonella-positive fecal dropping was 24 wk. In the daily Salmonella monitoring, only Salmonella Infantis was isolated from fecal droppings, in which the highest number of Salmonella Infantis organisms per fecal dropping was at 1 x 10(8) colony-forming units (cfu), while the lowest measured quantity was 1 x 10(3) cfu. It was noted that the frequency of Salmonella shedding in fecal droppings appeared to have a linear correlation (r = 0.85) with the number of Salmonella organisms (cfu) per fecal pellet (P < 0.05). Moreover, pulsed-field gel electrophoresis analysis of Salmonella Infantis isolates revealed a single identical pulsed-field pattern. Salmonella Enteritidis isolates from fecal droppings and internal organs also generated a single identical pulsed-field pattern. Interestingly, Salmonella Infantis was not isolated from any of the organs examined, while Salmonella Enteritidis was isolated from the spleen and liver of one roof rat. These results may indicate that wild roof rats could persistently carry Salmonella and contaminate commercial poultry facilities through intermittent fecal shedding. Moreover, Salmonella Enteritidis in wild roof rats appears to be more of a systemic infection, in which isolation is most likely to occur in internal organs, whereas Salmonella Infantis is more likely an enteric type of infection, in which isolation is most likely to occur in the intestinal contents. It is very plausible that layer chickens could become infected with Salmonella through ingestion of Salmonella-positive fecal droppings or feeds contaminated with these fecal droppings from infected resident roof rats. This is likely one of the major reasons why layer houses can be persistently infected by Salmonella even if the facilities are thoroughly cleaned and disinfected and if replacement stocks are obtained from Salmonella-free breeders and rearing units. It is therefore a noteworthy suggestion that rodent control programs inside poultry premises comprise an essential and effective tool in the management and control of Salmonella contamination in layer flocks.     

Biochemical and histological responses of Rattus novergicus (Wistar) infected by Echinostoma paraensei (Trematoda: Echinostomatidae)

Tests were performed to evaluate the biochemical alterations in Rattus norvegicus after infection by the intestinal trematode Echinostoma paraensei. The rodents received 150 metacercariae each, serum samples were collected and the parasite load was quantified weekly until the fifth week of infection. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions and hepatic glycogen were determined. All the animals exposed to the metacercariae were infected in the first week and worms were recovered up to the third week after infection. The levels of AST, ALT, GGT, bilirubin and globulin rose in the first and/or second week and declined thereafter to levels near those of the control group. In contrast, the level of total proteins in the plasma fell significantly in the first week while the ALKP activity went down only in the fourth and fifth weeks in relation to the control group. There was no significant difference in the levels of albumin, glycogen and glucose. Infection by E. paraensei in R. norvegicus causes changes in the hepatic function, possibly resulting from the cholestasis produced by the partial obstruction of the bile duct by the helminths.     

Pathological studies on central nervous tissues of rats infected with Rattus serotype hantavirus (SR-11 strain).

Newborn rats inoculated intraperitoneally with a hantavirus strain (SR-11) developed neurological signs such as ataxia and limb paralysis. The main histological lesions were scattered and multiple neuronal degeneration and necrosis with eosinophilic cytoplasmic inclusion bodies (CIB) in the brain, spinal cord and ganglia. Immunohistochemically, the viral antigen was detected in neurons, capillary endothelial and glial cells throughout the nervous tissue and CIB were identified as consisting of hantaviral antigen (nucleocapsid protein). Ultrastructurally, CIB in the neurons consisted of an accumulation of granular or filamentous materials, or both. They were seen near a well-developed Golgi apparatus and associated with well-developed rough endoplasmic reticulum and increased numbers of ribosomes. The present results suggested that this virus strain was highly infective in neurons of the newborn rats and that excess production of viral antigen which accumulated as CIB resulted in the neuronal changes.     

Isolation of Neospora caninum from naturally infected white-tailed deer (Odocoileus virginianus)

Attempts were made to isolate Neospora caninum from naturally infected white-tailed deer (Odocoileus virginianus). A total of 110 deer killed during the 2003 hunting season in Virginia region were used for the isolation of N. caninum. Of these, brains from 28 deer that had NAT titer of 1:200 were inoculated into interferon-gamma gene knock out (KO) mice. N. caninum was isolated from the tissues of three deer and all three isolates were mildly virulent to KO mice. Only one of the isolates could be adapted to in vitro growth. Protozoa in the tissues of KO mice reacted with N. caninum-specific polyclonal antibodies and N. caninum DNA was demonstrated in infected tissues by PCR assays; sequences of portions of the ITS-1 and gene 5 loci were identical to those in the public database. This is the first record of in vitro isolation of N. caninum from white-tailed deer and lends credence to the white-tailed deer as an intermediate host for this parasite.     

Quantification and morphometry of myenteric neurones in the jejunum of Holtzman rats (Rattus norvegicus)

With 2 figures and 3 tables SUMMARY: The morphological pattern of the myenteric plexus (MP) is species-specific, and little is known about this pattern in Holtzman rats. The aim of the current experiment was the morphological and quantitative study of myenteric neurones in the Holtzman rat jejunum. Hematoxylin-Eosin and NADH-diaphorase (NADH-dp) staining were used to assess muscular layer thickness, neurone cell body area (CBA) and nuclei area (NA). Muscular layer thickness was found to be 114.77 ± 14.89 μm. Neuronal densities across the subregions of the jejunum were similar: mesenteric, 11.78 ± 2.89/mm(2) ; intermediate, 12.06 ± 2.69/mm(2) ; and antimesenteric, 10.67 ± 1.89/mm(2) . As expected, there was positive correlation between the CBA and NA of 79.19, 79.26 and 78.5% in the mesenteric, intermediate and antimesenteric subregions of the jejunum, respectively. Medium-sized neurones predominated in the ganglionic arrangement of the MP. These results indicate that the NADH-dp myenteric neurones in the jejunum of Holtzman rats are similar in many aspects to those found in the ileum of Holtzman rats and to those found in the small intestine of Wistar rats, including their location, ganglionic disposition and predominance of medium-sized CBA. However, neuronal density in the jejunum is lower than in the ileum. Based on these results showing morphological similarities to the MP of the Wistar rat, the Holtzman strain can be used to investigate the effects of adverse conditions on the morphology of the MP.     

Isolation and identification of bubaline herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis) from Iran

In order to isolate buffaloes herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis), 16 buffalo heifers were selected from a herd. At first, animals were bled and their sera were tested by virus neutralization (VN) test, using bovine herpesvirus 1 (BoHV-1). According to the results of VN test and dexamethasone injection (0.1 mg/kg BW) for 5 consecutive days, the examined buffaloes were divided into 4 groups. Vaginal and nasal swabs were daily collected from all buffaloes from day 0 to 10 days later. Based on the cytopathic effects in cell culture, a herpesvirus was isolated only from nasal swabs of three seropositive buffaloes which they had received dexamethasone. The nasal swabs of these three buffaloes were also positive in PCR, using primers specific for ruminant herpesviruses gD gene. The identity of the isolated viruses was determined according to partial amino acid sequences of gD, deduced from the nucleotide sequences of the PCR products. On the basis of sequence alignment, phylogenetic analysis, and genetic distances, the three buffalo virus isolates were more closely related to BuHV-1 and BoHV-5 than to BoHV-1.

     

Contraceptive effect and potential side-effects of deslorelin acetate implants in rats (Rattus norvegicus): Preliminary observations

During the last ten years, numerous species have been treated with deslorelin implants to induce contraception. The aims of the study were 1) to assess contraceptive efficacy of 4.7 mg subcutaneous deslorelin implants in rats, 2) to determine the latency of contraceptive effect, and 3) to determine potential side effects. Three experimental females were implanted and their estrous cycle was studied by vaginal smear. Two weeks after implantation, a male whose fertility was previously assessed with a control female, was introduced into their cage. No female conceived during the 4 mo following implantation. Additionally, 38 pet rats were recruited from clients in practice to test for potential side effects, including 6 males and 32 females with a mean age of 14 mo. Local reaction and transient weight gain during the first 2 wk, as well as behavioral changes were recorded. According to this pilot study, deslorelin implant could be used as a contraceptive method in female rats. The latency period is about 2 wk. Nevertheless, it might be possible to refine the treatment further using hormonal measurements. The duration of contraceptive effect is to be determined in an upcoming study.     

Contracaecum sp. infection in Hoplias malabaricus (moncholo) from rivers and marshes of Colombia

Nematode infection indices were recorded in Hoplias malabaricus captured in six different rivers and a marsh belonging to the North Coast Basin of Colombia, and from the Amazon River, during February 2003-December 2004. Preliminary morphological analysis of nematodes indicated the presence of Contracaecum sp. Parasites were mostly found in the intestinal mesenteries and a very low percentage in muscle. Parasite prevalence in all sampling locations at the north of Colombia was 100%, whereas in the Amazon River it was 6.12%. The mean intensity in the different stations were as follows: Magdalena River at the City of Magangué (58.92+/-7.59), Magdalena river at the city of Zambrano (128.9+/-7.08), Sinú River (53.88+/-4.92), Dique Channel (207.3+/-59.52), Cauca River (77.26+/-9.35), Atrato River (21.11+/-2.6), San Jorge River (39.5+/-7.13), and Totumo Marsh (62.5+/-6.38). In average, all specimens of Hoplias malabaricus from the north coast basin of Colombia were infected with a mean intensity of 77.82+/-4.81 (1-466 parasites per host) whereas in fish from the Amazon River this value was significantly lower (intensity 1.0+/-0.0). Size and weight correlated significantly with parasite intensity in fish collected from sampling locations at the north of Colombia (R=0.240, P<0.001 and R=0.199, P=0.008, respectively). Moreover, a significant, but low and negative correlation was found between condition factor and parasite intensity (R=-0.159, P=0.034), suggesting a possible impact of parasites on fish health. These results suggest, for the first time, that the parasitism in Moncholo is a widespread phenomenon in Colombian rivers and could represent a risk factor for human consumers.