An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle,Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

Immunohistochemical analysis for G protein in the olfactory organs of soft-shelled turtle, Pelodiscus sinensis

In turtles, the epithelia lining the upper and lower chambers of the nasal cavity project axons to theventral and dorsal parts of the olfactory bulbs, respectively. In a semi-aquatic soft-shelled turtle,Pelodiscus sinensis, more than 1,000 odorant receptor genes have been found, but it is notknown where they are expressed. In this study, we aimed to clarify the distribution of cells expressing thesegenes in the olfactory organs of soft-shelled turtles. Immunoreactions for the Gαolf, the α subunit of Gprotein coupled to the odorant receptors, were detected on the surface of epithelia lining both the upper andlower chambers of the nasal cavity. The receptor cells in the epithelium of both chambers possessed cilia onthe tip of their dendrites, whereas microvillous, non-ciliated, receptor cells were not found. These datasuggest that the odorant receptor genes are expressed by the ciliated receptor cells in the upper and lowerchamber epithelia. Precise location of the vomeronasal epithelium is not known at present.     

Effects of acute cold exposure on oxidative balance and total antioxidant capacity in juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

Acute cold exposure may disturb the physiological homeostasis of the body in ectotherms. To date, there has been no information on the effects of cold exposure on homeostasis of reactive oxygen species (ROS) or antioxidant defense response in the Chinese soft‐shelled turtle, Pelodiscus sinensis. In this study, P. sinensis juveniles were acclimated at 28 °C, transferred to 8 °C as cold exposure for 12 h, then moved back to 28 °C rewarming for 24 h. We measured the ROS level and total antioxidant capacity (TAC) in the brain, liver, kidney and spleen at 2 and 12 h cold exposure, and at the end of the rewarming period. Malonaldehyde (MDA) and carbonyl protein were used as markers of oxidative damage. Turtles being maintained simultaneously at 28 °C were used as the control group. Cold exposure did not disturb the ROS balance in all 4 tissues, while rewarming raised the ROS level in the brain and kidney of P. sinensis. Cold exposure and rewarming decreased the TAC in the brain, liver and spleen but did not change the TAC in the kidney. MDA and carbonyl protein levels did not increase during the treatment, indicating no oxidative damage in all 4 tissues of P. sinensis. Our results indicated that extreme cold exposure did not impact the inner oxidative balance of P. sinensis, but more ROS was produced during rewarming. P. sinensis showed good tolerance to the harsh temperature change through effective protection of its antioxidant defense system to oxidative damage. This study provides basic data on the stress biology of P. sinensis.     

Bacteriological, serological, pathological and immunohistochemical studies of Mycoplasma bovis respiratory infection in veal calves and adult cattle at slaughter

Mycoplasma bovis is an important cause of calf pneumonia worldwide. In this study, we examined 140 cattle at slaughter comprising 70 veal calves and 70 beef cattle; 115 animals with pneumonic lesions and 25 without. Lung samples were submitted for bacteriological, histological, and M. bovis-immunohistochemical analyses. Serology for M. bovis was positive in 76% of beef cattle and 100% of veal calves. M. bovis was isolated only from veal calves in 16 out of 64 pneumonic cases. M. bovis was detected by immunohistochemistry in seven bacteriologically positive cases. M. bovis antigen was associated with bronchogenic necrosuppurative or fibrinonecrotizing lesions. Bacteriologically positive and immunohistochemical negative cases were associated with catarrhal bronchointerstitial pneumonia. Results suggest that M. bovis infection may develop into a severe necrosuppurative bronchopneumonia or fibrinonecrotizing pneumonia when associated with a high number of intralesional organisms or, conversely, into a mild catarrhal bronchointerstitial pneumonia when associated with a low number of organisms.     

Serologically defined Rhipicephalus (Boophilus) microplus larval antigens in BmLF3, a partially pure Sephacryl S-300 fraction of crude larval proteins

This report is designed to provide additional information regarding larval soluble proteins toward the planned development of a comprehensive database of Rhipicephalus (Boophilus) microplus proteins that elicit a humoral immune response in cattle as a result of natural ectoparasite infestation. Larval proteins of R. microplus are complex and the protein profile is not dominated by any major proteins. This report focuses upon an S-300 Sephacryl (molecular sieve) column fraction, fraction 3 (BmLF3). With the use of SDS-PAGE (without-2ME) and Western blotting with a composite pool of pre- and post-R. microplus larval infestation antiserum BmLF3 was found to contain 7 apparent common ixodid major antigens (207.3, 171.9, 98.0, 86.5, 65.7, 58.9, and 38.0 kDa), those potentially shared with other ixodid species, and 2 apparent R. microplus specific antigens evidenced by low-level antibody binding in crude BmLF3 (149.4 kDa) and HPLC peak 8 of BmLF3 (116.0 kDa). In addition, BmLF3 contains potent inhibitors of trypsin activity. However, these inhibitors of trypsin did not appear to elicit host antibodies as a result of natural ectoparasite exposure, as defined by Western blotting of reduced and denatured trypsin binding proteins purified by affinity chromatography.     

Serotype distribution and production of muramidase-released protein, extracellular factor and suilysin by field strains of Streptococcus suis isolated in the United States

Streptococcus suis is an important swine pathogen and a zoonotic agent. Differences in virulence have been noted among the 33 described serotypes, serotype 2 being considered the most virulent. In this study, we aimed at assessing the serotype distribution and the production of virulence-associated markers by strains recovered from diseased pigs in the United States (U.S.). Results showed that among the 100 strains evaluated, serotype 3 (20% of the isolates) and serotype 2 (17%) were the most prevalent. We then investigated the presence in these isolates of the genes sly, epf and mrp, encoding the virulence-associated markers suilysin (SLY), extracellular factor (EF) and muramidase-released (MRP) protein, respectively. The effective production of the markers by the strains was also verified. Results showed that the presence of the gene did not always correlate with actual expression of the respective protein. In the case of MRP, this was due, in most cases, to frameshift mutations at the 5′ end of the gene resulting in premature stop codons. The most prevalent phenotypes among U.S. strains were MRP⁺EF⁻SLY⁻ (40%) and MRP⁻EF⁻SLY⁺ (35%). Serotype distribution greatly differed from that reported in several European countries, as did the production of virulence markers, particularly for serotype 2. On the other hand, our results for the U.S. S. suis isolates are similar to those reported for Canadian strains, suggesting a common status in North America.     

救必应中药血清与抗菌药联合对产ESBLs细菌抑菌效果的研究

本试验主要探讨救必应中药血清与抗菌药联合诱导细菌传代的体外抑菌活性。制备救必应中药血清,采用二倍微量稀释法体外检测抗菌药的最小抑菌浓度(MIC),然后救必应中药血清与抗菌药联合诱导产超广谱β-内酰胺酶(extended spectrumβ-lactamases,ESBLs)细菌传代。救必应中药血清与抗菌药联合诱导细菌传代对耐药大肠杆菌有不同程度的抑菌活性。救必应中药血清可明显地增强β-内酰胺类抗菌药、氨基糖苷类抗菌药、喹诺酮类抗菌药和卡巴氧类抗菌药对耐药菌的抑制作用。     

Expression of Mycoplasma bovis variable surface membrane proteins in the respiratory tract of calves after experimental infection with a clonal variant of Mycoplasma bovis type strain PG45

The pathomorphological findings and the expression and distribution of variable surface protein antigens (Vsp) of Mycoplasma (M.) bovis were characterised immunohistochemically in lungs of eight calves following inoculation with a Vsp A-expressing clonal variant of M. bovis type strain PG45. Within 48 h post inoculation (p.i.) an innate immune response dominated by macrophages and neutrophils develops. The monoclonal antibodies (mAbs) 1A1 and 1E5 detected M. bovis Vsp antigens in paraffin tissue sections of seven calves. Vsp antigens were widely distributed and were already present at day two p.i. within macrophages and other lung compartments. Taken together, the results demonstrate that the bovine is unable to eliminate M. bovis during the time period examined. Based on the different immunohistochemical labelling patterns obtained with the mAbs, the results also support the speculation that the in vivo variability of Vsps together with immunological factors may contribute to the chronicity of pulmonary disease.     

Cross-sectional study of the seroprevalence to Borrelia burgdorferi sensu lato and granulocytic Ehrlichia spp. and demographic, clinical and tick-exposure factors in Swedish horses

A cross-sectional study of the seroprevalence to Borrelia burgdorferi sensu lato and granulocytic Ehrlichia spp. in Swedish horses was conducted to evaluate associations with demographic, clinical and tick-exposure factors. From September 1997–1998, blood samples from 2018 horses were collected from the animals presented to veterinary clinics affiliated with the Swedish Horserace Totalizator Board (regardless of the primary cause for consultation). Standardized questionnaires with information both from owners and attending veterinarians accompanied each blood sample. The apparent seroprevalences to B. burgdorferi s. l. and granulocytic Ehrlichia spp. were 16.8 and 16.7%, respectively. The northern region had the lowest seroprevalences. Four logistic models were developed (controlling for demographic variables). In the disease model of seropositivity to B. burgdorferi s. l., age, breed, geographic region, the serologic titer to granulocytic Ehrlichia spp., season and the diagnosis coffin-joint arthritis were significant. In the tick-exposure model of B. burgdorferi s. l., pasture access the previous year and gender were significant. Age, racing activity, geographic region, season and the serologic titer to B. burgdorferi s. l. were associated with positivity to granulocytic Ehrlichia spp. In the tick-exposure model of granulocytic Ehrlichia spp., pasture access was a risk factor. An interaction between racing activity and geographic region showed that the risk of positive serologic reactions to Ehrlichia spp. was increased in the horse population in the south and middle of Sweden, but only among horses not used for racing. Except for the positive association between coffin-joint arthritis and serologic reactions to B. burgdorferi s. l., there were no significant associations in the multivariable models between non-specific or specific clinical sign or disease with seropositivity to either of these agents.     

A retrospective study of the clinical hematology and the serum biochemistry tests made on canine dirofilariasis cases in an animal hospital population in Bangkok, Thailand

Heartworm disease, caused by the filarial nematode, Dirofilaria immitis, is a major, potentially life-threatening disease of dogs, with worldwide distribution and global significance. It is not only of veterinary importance but it also has zoonotic potential in many regions. It is considered as an endemic disease in Thailand, although clinical data about the disease is rarely reported. The objectives of this study were to characterize the clinical hematology and the biochemistry of canine dirofilariasis cases admitted to Chulalongkorn University, Small Animal Teaching Hospital in Bangkok, Thailand, from 2001-2003. All hematology and serum biochemistry parameter interpretations were based on reference values. A total of 1023 dogs were evaluated in this study. Dogs were divided into three groups based on their heartworm classification (microfilaremic, occult and negative). The major hematological findings in microfilaremic dogs were a mild to moderate anemia, mild to severe thrombocytopenia, marked leukocytosis, moderate to marked neutrophilia, eosinophilia and monocytosis. The most common serum biochemical abnormalities in microfilaremic dogs included increased alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase with a mean+/-SD of 311+/-299, 82+/-76 and 50+/-38 IU/L, respectively. It is likely that this parasitic infection led to the impairment of the hematological and biochemical status of the infected dogs. We believe that investigation of these laboratory based parameters, when associated with the clinical signs, is a very important approach to be considered in the routine clinical follow up, as well as being important for therapeutic evaluations.     

The effect of grain species, processing and time of feeding on the efficiency of feed utilization and microbial protein synthesis in sheep

This study investigated the effect of cereal grain species (sorghum, wheat, oats and barley), extent of processing (cracked barley, finely ground barley, and wet whole barley) and time of feeding (barley grain mixed with ryegrass hay or fed 2 h before hay was fed), on whole tract dry matter digestibility (WTDMD), and microbial protein synthesis (MPS), as a supplement to ryegrass hay when fed to rumen-cannulated sheep.Expected dry matter digestibility (EDMD) in mixtures of cereal grain and ryegrass hay was calculated by interpolation between in vitro dry matter digestibility (DMD) of each grain and the ryegrass hay. These were compared with measured actual WTDMD to detect positive or negative associative effects. Among grain species, the percentage difference in digestibility between actual WTDMD and EDMD was negative at − 6.6% units for wheat but positive at + 2.3%, + 4.3% and + 5.7% units for sorghum, oats, cracked or finely ground barley, respectively.As expected, the supplementation of sheep fed ryegrass hay with different sources of carbohydrates increased urinary allantoin output (as an indicator of MPS) when compared to sheep fed ryegrass hay alone. The concentration of urinary allantoin was significantly higher in sheep supplemented with sorghum (1916 mg/sheep/day) than wheat, oats or cracked barley ([mean ± S.E.M.] 1451 ± 24 mg/sheep/day) grain. There was a significantly higher urinary allantoin concentration in sheep fed cracked barley compared to finely ground barley or wet whole barley (1479 vs. 1095 vs. 1031 mg/sheep/day, respectively). There was no significant (P > 0.05) difference in urinary allantoin output, expressed as output/kg DM intake, when cracked barley was mixed and fed with the hay or fed 2 h before hay. However, in terms of total output of allantoin this was significantly higher (P < 0.05) (1479 vs. 1209 mg/sheep/day).In sacco degradability characteristics of organic matter and nitrogen for sorghum, oats, wheat, barley at different levels of processing and for ryegrass hay were also measured in the rumen of cannulated sheep.Among grain species, wheat had the highest effective organic matter degradability in the rumen (78.1%) while sorghum had the lowest. The effective degradability of protein of finely ground barley in the rumen was found to be higher than cracked barley or wet whole barley.Wheat grain, being highly degradable in the rumen, had a negative effect on WTDMD. In contrast, sorghum grain, being more slowly degradable in the rumen, would be expected to provide a substantially increased supply of energy to microbes over time in the rumen for MPS.Both cracked barley and finely ground barley also had a positive associative effect on WTDMD when fed with ryegrass increasing it by 5.7% units. The MPS was significantly higher (P < 0.05) in sheep fed cracked barley compared to finely ground barley or wet whole barley. This supports the hypothesis that slowly degrading carbohydrate sources synchronise more closely with available N from degradation of forage in the rumen.     

Changes in the risk management of Salmonella enterica subspecies diarizonae serovar 61:(k):1, 5, (7) in Swedish sheep herds and sheep meat due to the results of a prevalence study 2012

Background

The prevalence of Salmonella in food producing animals is very low in Sweden due to rigorous control programmes. However, no active surveillance is in place in sheep. The authorities decided to perform a prevalence study in sheep herds because findings at slaughter indicated that sheep associated S. diarizonae (S. enterica subspecies diarizonae serovar 61:(k):1, 5, (7)) might be common in sheep. Sampling was stratified by herd size in two groups, small herds with ≤ 30 animals and large herds with > 30 animals. In each stratum, 237 herds were selected at random. Faecal samples received from 244 out of the 474 randomly selected herds were analysed.

Results

A total of 40 of 100 (40%) of large herds and 17 of 144 (12%) of small herds were positive. The overall adjusted prevalence was 17.6% (95% CI, 12.9-22.2). Sheep associated S. diarizonae was detected in all counties (n = 21). Scientific opinions and an evaluation of on-farm control measures performed concluded that the impact of sheep associated S. diarizonae on human health is very low, and that risk management measures applied in response to findings of sheep associated S. diarizonae in sheep or sheep meat can be expected to have very little impact on reducing risks to human health. As a result, Swedish authorities decided to make an exemption for sheep associated Salmonella diarizonae in sheep and sheep meat in the current Salmonella control measures.

Conclusions

Sheep associated S. diarizonae is endemic in Swedish sheep herds. It is more common in large herds and not limited to certain parts of the country. The responsible authorities concluded that current risk management actions regarding sheep associated S. diarizonae in sheep and sheep meat are not proportional to the risk. This is the first time in the history of the Swedish Salmonella control programme that an exemption from the legislation has been made for a specific serovar. If there is any future indication of an increasing risk, due to e.g. change in the pathogenicity or development of antimicrobial resistance, the risk assessment will be re-evaluated and control measures reinforced if needed.