Further characterization of diabetes mellitus and body weight loss in males of the congenic mouse strain DDD.Cg-Ay

The A^y allele at the agouti locus causes obesity and promotes linear growth in mice. However, body weight gain stops between 16 and 17 weeks after birth, and then, body weight decreases gradually in DDD.Cg-A^y male mice. Body weight loss is a consequence of diabetes mellitus, which is genetically controlled mainly by a quantitative trait locus (QTL) on chromosome 4. This study aimed to further characterize diabetes mellitus and body weight loss in DDD.Cg-A^y males. The number of β-cells was markedly reduced, and plasma insulin levels were very low in the DDD.Cg-A^y males. Using a backcross progeny of DDD × (B6 × DDD.Cg-A^y) F₁-A^y, we identified one significant QTL for plasma insulin levels on distal chromosome 4, which was coincidental with QTL for hyperglycemia and lower body weight. The DDD allele was associated with decreased plasma insulin levels. When the DDD.Cg-A^y males were housed under three different housing conditions [group housing (4 or 5 DDD.Cg-A^y and DDD males), individual housing (single DDD.Cg-A^y male) and single male housing with females (single DDD.Cg-A^y male with DDD.Cg-A^y or DDD females)], diabetes mellitus and body weight loss were most severely expressed in individually housed mice. Thus, the severity of diabetes and body weight loss in the DDD.Cg-A^y males was strongly influenced by the housing conditions. These results demonstrate that both genetic and nongenetic environmental factors are involved in the development of diabetes mellitus and body weight loss in the DDD.Cg-A^y males.     

Distribution of the sex chromosome during mouse spermatogenesis in testis tissue sections

During mammalian spermatogenesis, spermatogenic cells undergo mitotic division and are subsequently divided into haploid spermatids by meiotic division, but the dynamics of sex chromosomes during spermatogenesis are unclear in vivo. To gain insight into the distribution of sex chromosomes in the testis, we examined the localization of sex chromosomes before and after meiosis in mouse testis sections. Here, we developed a method of fluorescence in situ hybridization (FISH) using specific probes for the X and Y chromosomes to obtain their positional information in histological testis sections. FISH analysis revealed the sex chromosomal position during spermatogenesis in each stage of seminiferous epithelia and in each spermatogenic cell. In the spermatogonia and leptotene spermatocytes, sex chromosomes were distantly positioned in the cell. In the zygotene and pachytene spermatocytes at prophase I, X and Y chromosomes had a random distribution. After meiosis, the X and Y spermatids were random in every seminiferous epithelium. We also detected aneuploidy of sex chromosomes in spermatogenic cells using our developed FISH analysis. Our results provide further insight into the distribution of sex chromosomes during spermatogenesis, which could help to elucidate a specific difference between X and Y spermatids and sex chromosome-specific behavior.     

Evaluation of the oral 13C-bicarbonate technique for measurements of energy expenditure in dogs before and after body weight reduction

Background

Overweight and obesity are the most common nutritional disorders in dogs and may lead to various secondary diseases and decreased lifespan. In obesity research, measurement of energy expenditure (EE) and determination of the energy requirements are essential. The objective with this study was to validate and evaluate the suitability of the oral ¹³C-bicarbonate technique (o¹³CBT) for measuring EE in dog obesity studies. A further objective was to investigate the impact of body weight (BW) reduction and changes in body composition on the EE when measured under conditions corresponding to the basal metabolic rate (BMR).

Results

The EE in five privately owned, overweight dogs was measured simultaneously with the o¹³CBT and indirect calorimetry (IC) for comparison of the results. Two measurements per dog were performed under the same standardised conditions (i.e. fasted and resting state) at the start, and after completing a 12-week BW reduction program. Additionally, measurements of body composition by Dual-energy X-ray absorptiometry (DEXA) were conducted at the beginning and at the end of the BW reduction program. There were no differences in EE results obtained by the o¹³CBT and IC. Overweight and the BW reduction did not affect the estimates for the respiratory quotient (RQ) or the recovery factor for the ¹³C-tracer (RF), both needed when using the o¹³CBT. The dogs lost 16% (SD ± 2.0) of their initial BW in reduced fat mass (P < 0.001), whereas fat free mass (FFM) remained unchanged. There was no effect of the BW reduction on the determined EE expressed in kJ/kg BW/d, or in kJ/kg BW^0.75/d. However, EE was lower (P < 0.001) after the BW reduction program when expressed in relation to FFM (kJ/kg FFM/d).

Conclusions

Results from the present study show that the o¹³CBT can be a used in obesity research to determine EE in fasted dogs and under resting conditions. Furthermore, the results suggest that the BMR does not change with reduced BW in overweight dogs as long as the FFM remains unchanged. This indicates that the BMR to maintain one gram of fat is equal to maintaining one gram of FFM in overweight dogs.     

Tissue-specific expression of the NOD-like receptor protein 3 in BALB/c mice

Activation of the innate immune system requires recognition of pathogen-associated molecular patterns, such as NOD-like receptors. The NOD-like receptor protein 3 (NLRP3) inflammasome is involved in induction of the pro-inflammatory cytokine, IL-1β, and subsequent inflammatory responses. NLRP3 inflammasome plays important roles in the inflammatory and innate immune responses associated with autoimmune/inflammatory syndrome. However, analysis of the tissue distribution and expression profiles in BALB/c mice is still incomplete. In this study, we investigated the tissue distribution and expression pattern of NLRP3 in BALB/c mice to further elucidate its function in innate immunity in this commonly used laboratory animal model. NLRP3 mRNA expression levels and tissue distribution of the protein were investigated by real-time quantitative PCR and immunohistochemical analyses, respectively. NLRP3 mRNA expression was higher in the kidney and inguinal lymph nodes than in other tissues. Cytoplasmic expression of NLRP3 was detected in the epithelial reticular cells of the spleen and thymus, lymphocytes in the inguinal lymph nodes, cardiac muscle cells, cerebral cortex neurons, alveolar macrophages, renal tubule cells and liver sinusoidal endothelial cells. The results of this study will assist investigators in interpreting site-specific functions and roles of NLRP3 in inflammatory responses.     

Helicobacter apodemus sp. nov., a new Helicobacter species identified from the gastrointestinal tract of striped field mice in Korea

A novel Helicobacter species was identified from the gastrointestinal tract of the Korean striped field mouse (Apodemus agrarius). Biochemical testing, ultrastructure characterization, and 16S rRNA gene sequence analysis suggested that this bacterium represents a distinct taxon. The bacterium was positive for urease activity, susceptible to cephalothin and nalidixic acid, and weakly positive for oxidase and catalase activity. Electron microscopy revealed that the bacterium has spirally curved rod morphology with singular bipolar nonsheathed flagella. Genotypically, the isolated bacterial strains (YMRC 000215, YMRC 000216, and YMRC 000419) were most closely related to a reference strain of Helicobacter mesocricetorum (97.25%, 97.32%, and 97.03% 16S rRNA sequence similarities, respectively). The 16S rRNA sequences of these strains were deposited into GenBank under accession numbers {"type":"entrez-nucleotide","attrs":{"text":"AF284754","term_id":"18389594","term_text":"AF284754"}}AF284754, {"type":"entrez-nucleotide","attrs":{"text":"AY009129","term_id":"18409527","term_text":"AY009129"}}AY009129, and {"type":"entrez-nucleotide","attrs":{"text":"AY009130","term_id":"18409528","term_text":"AY009130"}}AY009130, respectively. We propose the name Helicobacter apodemus for this novel species.     

Genetic variability of the prion protein gene (PRNP) in wild ruminants from Italy and Scotland

The genetics of the prion protein gene (PRNP) play a crucial role in determining the relative susceptibility to transmissible spongiform encephalopathies (TSEs) in several mammalian species. To determine the PRNP gene variability in European red deer (Cervus elaphus), roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra), the PRNP open reading frame from 715 samples was analysed to reveal a total of ten single nucleotide polymorphisms (SNPs). In red deer, SNPs were found in codons 15, 21, 59, 78, 79, 98, 136, 168 and 226. These polymorphisms give rise to 12 haplotypes, and one of which is identical to the PRNP of American wapiti (Rocky Mountain elk, Cervus elaphus nelsoni). One silent mutation at codon 119 was detected in chamois and no SNPs were found in roe deer. This analysis confirmed that European wild ruminants have a PRNP genetic background that is compatible with TSE susceptibility, including chronic wasting disease.     

The radioprotective effects of the hexane and ethyl acetate extracts of Callophyllis japonica in mice that undergo whole body irradiation

The radioprotective activity of extracts from the red seaweed Callophyllis (C.) japonica was investigated in mice that underwent whole-body exposure to gamma radiation. A methanol extract of C. japonica and its fractions [hexane, ethyl acetate (EtOAc), butanol and the remaining H₂O] were used. Each fraction (100 mg/kg body weight) was administered intraperitoneally (i.p.) 2 times into the BALB/c mice, once at 1 and once at 24 h before exposure to 9 Gray (Gy) of gamma radiation. Pre-irradiation administration of the hexane and EtOAc fractions saved the mice, with their survival rates being greater than 80% at 30 days post-irradiation; the mice that were pretreated with the other fractions showed survival rates lower than 20% over the same time period. To examine the effect of each C. japonica fraction on the survival of intestinal and bone marrow stem cells, the number of intestinal crypts and bone marrow cells in the gamma-irradiated mice were examined. Pre-treatment of mice (i.p., 100 mg/kg body weight at 1 and 24 h before irradiation) with the hexane or EtOAc fraction prior to 6-Gy irradiation significantly protected the number of jejunal crypts and bone marrow cells at 9 days after irradiation. These findings suggest that certain extracts from C. japonica, when they are administered prior to irradiation, play an important role in the survival of irradiated mice, and this is possibly due to the extracts protecting the hematopoietic cells and intestinal stem cells against gamma irradiation.     

麻叶荨麻醇提液对大鼠离体子宫运动的影响

为探究荨麻对子宫运动的影响及其可能机制,本试验用生物信号处理系统测定了浓度递增(0.5%、1.0%、2.0%、4.0%、8.0%和16.0%)的麻叶荨麻(Urtica cannabina,UC)醇提液对未孕大鼠离体子宫收缩频率、收缩张力和收缩幅度的影响,并用特异性M-受体阻断剂硫酸阿托品(atropine,A,2.5 μg/mL)和2.0% UC联合用药,分为3个处理: UC单独处理、UC+A和A+UC,观察UC对子宫收缩的影响是否与M-受体有关.结果显示,在UC浓度递增到4.0%时收缩频率较用药前显著增加(P <0.05);浓度递增到4.0%、8.0%时,收缩幅度较用药前极显著降低(P <0.01);但浓度递增UC对子宫收缩张力影响不显著(P >0.05).一次性加入2.0% UC后子宫收缩张力迅速增加并接近僵直,之后逐渐降低.无论UC+A或A+UC处理都使子宫收缩频率较用药前明显加快,收缩张力极显著增加(P <0.01),但2个联合用药处理组之间差异不显著(P >0.05);UC+A和A+UC处理组收缩张力较UC单独用药组均极显著降低(P <0.01).结果表明,UC可兴奋未孕大鼠离体子宫,A可缓解UC引起的子宫僵直或张力增加,但不能阻断UC引起的子宫收缩频率增加.提示UC对未孕大鼠离体子宫的兴奋作用和激动与M-受体有一定关系,但可能还存在其他机制.     

A study on the effect of different rifle calibres in euthanisation of grey seals (Halichoerus grypus) in seal traps in the Baltic Sea

Background

In recent years, the euthanasia of seals has been discussed internationally and concern has been raised regarding the use of rifles, the effect of different calibres, and which calibres are sufficient for humane euthanasia. This study therefore investigated the effect of different firearm calibres on euthanasia of grey seals (Halichoerus grypus) in traps, and provides information for the development and refinement of regulations for hunting seals in the wild.

Findings

The effect of different calibres was studied in 19 seals shot in the head and neck at close range. All seals were necropsied and radiographed to characterize the injuries caused by the bullets. All tested calibres, 5.6 mm bullet diameter or larger, and .12 shotgun, were sufficiently effective to cause severe skull fractures, meningeal haemorrhages and instant death.

Conclusions

Rifles with 5.6 mm bullet diameter or larger, and a .12 shotgun loaded with a slug fired at close range to the head and neck of grey seals all caused instant death and can therefore be recommended for hunting seals in the wild.     

Protective Effect of Stachybotrys microspora Triprenyl Phenol-7on the Deposition of IgA to the Glomerular Mesangium in Nivalenol-induced IgA Nephropathy Using BALB/c Mice

Activators of tissue proteolysis including Stachybotrys microspora triprenyl phenol (SMTP)-7 are a new class of agents that are expected to be effective for amelioration of chronic tissue destructive diseases. The present study was performed to examine whether SMTP-7 is effective for the amelioration or protection of early-stage IgA nephropathy (IgAN) induced by nivalenol (NIV) in female BALB/c mice. In Experiment 1, mice were administered NIV at 24 ppm in diet for 8 weeks, and during the NIV treatment, they were intraperitoneally injected with SMTP-7 (10 mg/kg) three times a week. In Experiment 2, mice were injected similarly with SMTP-7 during the last 4 weeks of a 16-week NIV treatment. Immunofluorescence analysis revealed an inhibitory effect of SMTP-7 on the glomerular deposition of IgA in Experiment 1; however, it was ineffective in Experiment 2. On the other hand, SMTP-7 did not affect the serum concentration of IgA in both experiments. These results suggest that SMTP-7 has a potential to decrease the progression of IgAN induced by NIV through inhibition of local accumulation of IgA in the glomerular mesangium, while it was ineffective for suppression of IgA production. On the other hand, SMTP-7 was found to be ineffective for already deposited IgA, suggesting that SMTP-7 may not be effective for ameliorating advanced IgAN.     

Abattoir-based study on the epidemiology of caprine tuberculosis in Ethiopia using conventional and molecular tools

Background

Despite the important role of goats for meat and milk production in Ethiopia, little information is available on the epidemiology of caprine tuberculosis (TB). Caprine TB is important as milk is usually consumed raw particularly by Ethiopian pastoralists. The objectives of the present study were to estimate the prevalence of TB in goats at an abattoir, to evaluate associated risk factors and to characterize the causative mycobacteria.

Methods

A cross-sectional study was conducted on 1990 randomly selected male goats that were slaughtered at Luna Export Abattoir of central Ethiopia. Postmortem examination, mycobacterial culturing and molecular typing techniques like genus typing, deletion typing and spoligotyping were used.

Result

The overall prevalence of caprine TB-like lesions was 3.5%. The lesion prevalence increased significantly with increasing age. Mycobacteria were found by culture and seen as acid fast bacilli in 12% of the goats with TB-like lesions. Characterization of the eight isolates using multiplex polymerase chain reaction (PCR) indicated that five of them belonged to the genus Mycobacterium. Four of the latter were confirmed to be members of the M. tuberculosis complex. Further characterization of the three M. tuberculosis isolates by spoligotyping identified them as type SIT53 and two new spoligotypes.

Conclusion

The isolation of M. tuberculosis from goats in this study indicates a potential risk of transmission of M. tuberculosis between humans and goats.     

Efficiency of Matricaria chamomilla CH12 and number of doses of rabies vaccine on the humoral immune response in cattle

This study evaluated the effect of Matricaria chamomilla and vaccination frequency on cattle immunization against rabies. Four groups (n = 15 /group) were treated with or without Matricaria chamomilla CH₁₂ and vaccinated with one or two doses of rabies vaccine (30 day interval). No effect of chamomile was found on cattle immunization against rabies; however, antibody titers were protective in cattle vaccinated twice, while 93.3% of cattle vaccinated only once had titers under 0.5 UI/ml after 60 days. In conclusion, the use of chamomile did not alter the humoral immune response in cattle, and two vaccine doses are suggested for achieving protective antibody titers.     

Preliminary Studies on the Concentration of Na+,K+-ATPase in Skeletal Muscle of Draught Cattle in Mozambique: Effect of Sex,Age and Training

The effect of training on the potential for work in draught cattle was assessed by measuring the Na⁺,K⁺-ATPase in the muscle cell membrane and the elevation in the concentration of K⁺ in plasma during exercise. Biopsies of the semitendinosus muscle and venous blood samples were taken from the cattle used for draught work in Mozambique. No differences were found in the plasma ion or Na⁺,K⁺-ATPase concentrations in samples taken from Nguni, Africander and Angoni breeds. There were no significant differences in plasma ions (Na⁺, K⁺ and Cl^–) or muscle Na⁺,K⁺-ATPase concentrations between the Angoni males and females, although the males showed an increase in Na⁺,K⁺-ATPase with age, while the females showed a decrease. The increase in males might be attributed to their higher level of activity in the herds than that of females. After a training period of 15 days, a significant increase in Na⁺,K⁺-ATPase concentration in semitendinosus muscle was found in Angoni cattle. In females, this was significant after 8 days of training (about 30%); in males after 15 days of training (about 16%). On day 15, there was a reduction in the elevation of plasma K⁺ during the 2 h of draught work, indicating an increased capacity of the Na⁺,K⁺ pumps to maintain the extracellular K⁺ concentration in working muscles and a possible delay in the moment of fatigue.     

Investigations on the efficacy of routinely used phenotypic methods compared to genotypic approaches for the identification of staphylococcal species isolated from companion animals in Irish veterinary hospitals

Background

Identification of Staphylococci to species level in veterinary microbiology is important to inform therapeutic intervention and management. We report on the efficacy of three routinely used commercial phenotypic methods for staphylococcal species identification, namely API Staph 32 (bioMérieux), RapID (Remel) and Staph-Zym (Rosco Diagnostica) compared to genotyping as a reference method to identify 52 staphylococcal clinical isolates (23 coagulase positive; 29 coagulase negative) from companion animals in Irish veterinary hospitals.

Results

Genotyping of a 412 bp fragment of the staphylococcal tuf gene and coagulase testing were carried out on all 52 veterinary samples along with 7 reference strains. In addition, genotyping of the staphylococcal rpoB gene, as well as PCR-RFLP of the pta gene, were performed to definitively identify members of the Staphylococcus intermedius group (SIG). The API Staph 32 correctly identified all S. aureus isolates (11/11), 83% (10/12) of the SIG species, and 66% (19/29) of the coagulase negative species. RapID and Staph-Zym correctly identified 61% (14/23) and 0% (0/23) respectively of the coagulase-positives, and 10% (3/29) and 3% (1/29) respectively of the coagulase-negative species.

Conclusions

Commercially available phenotypic species identification tests are inadequate for the correct identification of both coagulase negative and coagulase positive staphylococcal species from companion animals. Genotyping using the tuf gene sequence is superior to phenotyping for identification of staphylococcal species of animal origin. However, use of PCR-RFLP of pta gene or rpoB sequencing is recommended as a confirmatory method for discriminating between SIG isolates.     

Effect of dietary fiber on methane production in Chinese Lantang gilts

The objective of this study was to determine the effect of dietary fiber on methane (CH₄) production in pigs using the Chinese native Lantang gilts as study model. The study consisted of two experiments. In the first, 12 Lantang gilts (58.7±0.37 kg), individually housed in open-circuit respiration chambers were randomly divided into two groups (six replicates per dietary group) and fed either with low fiber diet [LFD; neutral detergent fiber (NDF)=201.5 g/kg] or high fiber diet (HFD; NDF=329.7 g/kg). Wheat bran was the main source of fiber for the LFD while ground rice hull (mixture of rice bran and rice hull) was used in the HFD. Results of the study showed that gilts fed LFD recorded higher (P<0.05) digestibility coefficients for dry matter (DM), total organic carbon (TOC), acid detergent fiber (ADF) and cellulose than those in the HFD. However, digestibility coefficient for NDF did not differ between treatments but that for hemicellulose was higher for HDF than for LDF. Because of the higher NDF and hemicellulose contents in the diet, pigs in the HFD recorded higher (P<0.05) amount of digested NDF (126.1 vs. 83.6 g/d) and hemicellulose (38.7 vs. 11.9 g/d) than those fed LFD. The higher amount of digested NDF and hemicellulose recorded for the HFD treatment was inconsistent with the lower (P<0.01) daily CH₄ production from gilts fed the HFD compared to that fed LFD (2.46 vs. 3.90 L/pig). To better explain for the unexpected higher CH₄ production from pigs fed LFD, an in vitro fermentation study was conducted using a factorial design comprised of two inocula (collected from low fiber and high fiber diets) and eight substrates (LFD, HFD, wheat bran, ground rice hull and their respective NDF residues). Results of the in vitro trial seems to suggest that the low CH₄ production from the HFD animals was primarily the results of low fermentation rates of HFD and the ground rice hull. However, the calculated CH₄ productions based on the molar percentage of volatile fatty acids (VFA) in both, in vivo and in vitro experiments reaffirmed the in vivo result, that is, CH₄ production from pigs fed HFD was lower than that fed LFD. Although there is a lack of scientific data from this study to fully explain for the unexpected lower CH₄ production from pigs fed the HFD, our results seems to suggest that quantity of digested fiber (including NDF) was not the sole factor affecting enteric CH₄ production in pigs.     

The Effect of Treatment with a 1% Injectable Formulation of Moxidectin during the Rainy Season and at the Beginning of the Dry Season on Gastrointestinal Nematodes in Cattle from Communal Areas in Zimbabwe

The effect of 1% moxidectin/cydectin at 0.2 mg/kg live weight on gastrointestinal nematodes and on the growth of calves, weaners and cows was investigated in five communal areas on the highveld of Zimbabwe. Three field experiments were carried out between March 1996 and June 1997. In experiment 1, treatment was administered in all five areas at the end of the rainy season in March 1996, followed by a further treatment at the beginning of the dry season in May/June 1996. In experiment 2, the treatment was administered in three areas at the end of the rainy season in March 1997. In experiment 3, treatment was administered in one area at the beginning of the dry season in April 1997. Large numbers of eggs were present in the faeces of calves and weaners at the start of experiments 1 and 2. Epg values were lower in cows and in all age categories in experiment 3. There was a statistically significant reduction in epg values in calves, weaners and cows following treatment with a reduction of 90–99% in all cases except in cows in experiment 3, where no meaningful assessment was possible owing to the low egg counts in both the treated and control cows. The dominating larval types in faecal cultures were Cooperia and Haemonchus. Trichostrongylus, Oesophagostomum and Bunostomum were also found. Following treatment, Haemonchus was suppressed far more than Cooperia. This may be related to a longer residual effect against abomasal parasites like Haemonchus in comparison to small intestinal worms like Cooperia. Anthelmintic treatment conferred significant weight gain advantages (p<0.05) on treated calves, weaners and cows. The weight gains are discussed in relation to disease and nutrition.