Visual landmark‐directed scatter‐hoarding of Siberian chipmunks Tamias sibiricus

Spatial memory of cached food items plays an important role in cache recovery by scatter‐hoarding animals. However, whether scatter‐hoarding animals intentionally select cache sites with respect to visual landmarks in the environment and then rely on them to recover their cached seeds for later use has not been extensively explored. Furthermore, there is a lack of evidence on whether there are sex differences in visual landmark‐based food‐hoarding behaviors in small rodents even though male and female animals exhibit different spatial abilities. In the present study, we used a scatter‐hoarding animal, the Siberian chipmunk, Tamias sibiricus to explore these questions in semi‐natural enclosures. Our results showed that T. sibiricus preferred to establish caches in the shallow pits labeled with visual landmarks (branches of Pinus sylvestris, leaves of Athyrium brevifrons and PVC tubes). In addition, visual landmarks of P. sylvestris facilitated cache recovery by T. sibiricus. We also found significant sex differences in visual landmark‐based food‐hoarding strategies in Siberian chipmunks. Males, rather than females, chipmunks tended to establish their caches with respect to the visual landmarks. Our studies show that T. sibiricus rely on visual landmarks to establish and recover their caches, and that sex differences exist in visual landmark‐based food hoarding in Siberian chipmunks.     

Selenium‐enriched Saccharomyces cerevisiae improves growth,antioxidant status and selenoprotein gene expression in Arbor Acres broilers

One hundred and fifty 7‐day‐old Arbor Acres broilers were randomly assigned into five groups: group 1 served as a control that was fed a basal diet without selenium (Se) supplementation; groups 2, 3 and 4 were fed the basal diet supplemented with 0.15, 0.5 and 1.5 mg Se as Se‐enriched Saccharomyces cerevisiae (SSC) per kg of diet; and group 5 was fed the basal diet supplemented with 0.15 mg per kg of Se as sodium selenite (SS). Growth performance, glutathione peroxidase (GP_X) and superoxide dismutase (SOD) activities, total antioxidant capacity (T‐AOC), and malondialdehyde (MDA) content in plasma and liver, and cellular glutathione peroxidase (GP_X‐1) and phospholipid hydroperoxide glutathione peroxidase (GP_X‐4) mRNA levels in liver were determined. Compared with group 1, groups 2–4 exhibited higher body weights (p < 0.05), lower feed/gain ratios, and higher GP_X activities in plasma (p < 0.05) and GP_X and SOD activities and GP_X‐1 and GP_X‐4 mRNA levels in liver (p < 0.05). Compared with group 5, group 2 exhibited higher GP_X activity in plasma on day 21 (p < 0.05). Compared with group 2 and 5, group 3 exhibited lower MDA content in plasma on day 7 (p < 0.05), higher GP_X activity in plasma, SOD activity and GP_X‐1 mRNA levels in liver on day 14 and 21 (p < 0.05), and higher GP_X‐4 mRNA levels on day 14 (p < 0.05). Compared with group 4, group 3 exhibited lower MDA contents in plasma on day 14 (p < 0.05) and in liver on day 21 (p < 0.05), higher T‐AOC in plasma and higher GP_X‐1 mRNA levels on day 14 and 21 (p < 0.05), and higher SOD activity in plasma and higher SOD and GP_X activities in liver on day 21 (p < 0.05). Thus, SSC improves growth and antioxidant status of broilers; the short‐term bioavailability of SS was faster than that of SSC, but the long‐term bioavailability of SSC was greater than SS.     

Fine mapping the number of corpora lutea quantitative trait loci on SSC3: Analysis of the porcine follicle‐stimulating hormone receptor gene

In females, follicle‐stimulating hormone (FSH) targets a FSH receptor (FSHR) expressed only on granulose cells, inducing maturation of the ovarian follicles. We hypothesized that genetic variants in the FSHR gene influence litter size by affecting the number of corpora lutea. We fine‐mapped a region of Sus Scrofa chromosome 3 that contains quantitative trait loci for corpora lutea. Polymorphisms were detected in the exons and 5′ flanking region of the porcine FSHR gene, a positional candidate for the statistically most significant of the quantitative trait loci. Finally, 248 F₂ animals from a Duroc and Meishan cross were genotyped for three FSHR SNPs at positions 74, 532 and 1166, and these were correlated with the phenotypes of litter size and corpus luteum number. Three haplotypes were identified: M1 (G/G/C), M2 (C/A/T) and D (C/A/C). In the F₂ population, the M1 haplotype was associated with a greater number of corpora lutea (P < 0.01) and also seemed to be associated with increased litter size, although the association was not significant (P = 0.2571). Some polymorphisms resulting in amino acid substitutions in these genes were excluded from the polymorphisms possibly responsible for the number of corpora lutea.     

The effect of 25‐hydroxyvitamin D3 and phytase inclusion on pig performance,bone parameters and pork quality in finisher pigs

The objective of this study was to investigate the effects of supplementing both phytase and 25‐hydroxyvitamin D₃ (25‐OH ‐D?) on pig performance, nutrient digestibility, carcass characteristics, bone parameters and pork quality in finisher pigs. The experimental design was a 2 × 2 factorial comprising of four dietary treatments. One hundred and twenty pigs (60 male, 60 female) were blocked according to live weight and sex and allocated to the following dietary treatments: low P (4.81 g/kg) diet (basal) (T1); low P diet + phytase (T2); low P diet + 25‐OH ‐D? (T3) and low P diet + phytase + 25‐OH ‐D? (T4). Pigs supplemented with phytase had a lower average daily feed intake (ADFI ) (2.45 kg vs. 2.59 kg; p  < 0.05) and lower feed conversion ratio (FCR ) (2.74 kg/kg vs. 2.85 kg/kg; p  <  0.05) compared to pigs offered the nonphytase diets. Pigs offered phytase diets had a higher (p  <  0.05) coefficient of apparent total tract digestibility (CATTD ) of ash, phosphorous (P) and calcium (Ca) compared with pigs offered the nonphytase supplemented diets. Pigs offered the 25‐OH ‐D₃ diets had a higher CATTD of N and ash. Pigs offered the phytase diets had increased (p  <  0.05) bone DM , ash, Ca, P and density compared to the nonphytase diets. There was a significant interaction (p  <  0.05) between phytase and 25‐OH ‐D₃ on cook loss. Pigs offered 25‐OH ‐D₃ had increased cook loss over the basal diet; however, there was no effect on cook loss when phytase and 25‐OH ‐D₃ were offered in combination compared to the phytase only diet. Pigs offered 25‐OH ‐D₃ exhibited higher (p  <  0.05) Warner Bratzler shear force values and lower (p  <  0.05) pork lightness (L *) surface colorimeter values. In conclusion, there was no benefit to offering a combination of phytase and 25‐OH ‐D₃ on pig performance, bone parameters or pork quality.     

Sex ratio variation and sex determination in the mallee dragon Ctenophorus fordi

Recent evidence suggests that many Australian agamids show temperature‐dependent sex determination (TSD) with variation in sex determining mechanisms among closely related taxa. However, as shown in other vertebrates, sex ratios can also be influenced by genetic or phenotypic differences among females in their propensity to produce sons or daughters, and these influences might confound any thermal effects of incubation per se. To address these issues, we investigated the determinants of sex ratios in the mallee dragon Ctenophorus fordi, together with a detailed analysis of karyotypes. There was no detectable variation in sex ratios arising from variation among females, clutches or incubation temperatures, which might indicate genetic sex determination for this species. However, there was no evidence of cytologically distinct sex chromosomes using standard banding techniques. The sex ratio pattern in C. fordi strongly contrasts with the results for the congener Ctenophorus pictus, where sex ratios show variation among females. Thus, Australian agamids offer promising opportunities to address fundamental issues in sex ratio biology.     

Enhancement of viability of a probiotic Lactobacillus strain for poultry during freeze‐drying and storage using the response surface methodology

A rotatable central composite design (CCD) was used to study the effect of cryoprotectants (skim milk, sucrose and lactose) on the survival rate of a probiotic Lactobacillus strain, L. reuteri C10, for poultry, during freeze‐drying and storage. Using response surface methodology, a quadratic polynomial equation was obtained for response value by multiple regression analyses: Y = 8.59546 ? 0.01038 X₁ ? 0.09382 X₂ ? 0.07771 X₃ ? 0.054861 X₁² ? 0.04603X₃² ? 0.10938 X₁X₂. Based on the model predicted, sucrose exerted the strongest effect on the survival rate. At various combinations of cryoprotectants, the viability loss of the cells after freeze‐drying was reduced from 1.65 log colony forming units (CFU)/mL to 0.26–0.66 log CFU/mL. The estimated optimum combination for enhancing the survival rate of L. reuteri C10 was 19.5% skim milk, 1% sucrose and 9% lactose. Verification experiments confirmed the validity of the predicted model. The storage life of freeze‐dried L. reuteri C10 was markedly improved when cryoprotectants were used. At optimum combination of the cryoprotectants, the survival rates of freeze‐dried L. reuteri C10 stored at 4°C and 30°C for 6 months were 96.4% and 73.8%, respectively. Total viability loss of cells which were not protected by cryoprotectants occurred after 12 and 8 weeks of storage at 4°C and 30°C, respectively.     

Genome-wide linkage and QTL mapping in porcine F2 families generated from Pietrain, Meishan and Wild Boar crosses

Three informative pig F₂ families based on European Wild Boar (W), Meishan (M) and Pietrain (P) crosses have been used for genome‐wide linkage and quantitative trait loci (QTL) analysis. Altogether 129 microsatellites, 56 type I loci and 46 trait definitions (specific to growth, fattening, fat deposition, muscling, meat quality, stress resistance and body conformation) were included in the study. In the linkage maps of M × P, W × P and W × M families, average spacing of markers were 18.4, 19.7 and 18.8 cM, the numbers of informative meioses were 582, 534 and 625, and the total lengths of autosomes measured were 27.3, 26.0 and 26.2 Morgan units, respectively. Maternal maps were on average 1.3 times longer than paternal maps. QTLs contributing more than 3% of F₂ phenotypic variance could be identified at p < 0.05 chromosome‐wide level. Differences in the numbers and positions of QTLs were observed between families. Genome‐wide significant QTL effects were mapped for growth and fattening traits on eight chromosomes (1, 2, 4, 13, 14, 17, 18 and X), for fat deposition traits on seven chromosomes (1, 2, 3, 4, 6, 7 and X), for muscling traits on 11 chromosomes (1, 2, 3, 4, 6, 7, 8, 12, 14, 15 and X), for meat quality and stress resistance traits on seven chromosomes (2, 3, 6, 13, 16, 18 and X), and QTLs for body‐conformation traits were detected on 14 chromosomes. Closely correlated traits showed similar QTL profiles within families. Major QTL effects for meat quality and stress resistance traits were found on SSC6 in the interval RYR1‐A1BG in the W × P and M × P families, and could be attributed to segregation of the RYR1 allele T derived from Pietrain, whereas no effect in the corresponding SSC6 interval was found in family W × M, where Wild Boar and Meishan both contributed the RYR1 allele C. QTL positions were mostly similar in two of the three families for body conformation traits and for growth, fattening, fat deposition and muscling traits, especially on SSC4 (interval SW1073‐NGFB). QTLs with large effects were also mapped on SSC7 in the major histocompatibility complex (MHC) (interval CYP21A2‐S0102) and affected body length, weight of head and many other traits. The identification of DNA variants in genes causative for the QTLs requires further fine mapping of QTL intervals and a positional cloning. However, for these subsequent steps, the genome‐wide QTL mapping in F₂ families represents an essential starting point and is therefore significant for animal breeding.     

Polymorphism in 5′ untranslated region of heat‐shock protein 70 gene as marker of post‐partum anoestrus in Murrah buffaloes

The enormous production potential of buffaloes has never been accomplished due to various reproductive insufficiencies. Among them, post‐partum anoestrus, a multifactorial disorder, is predominant but any genetic association is yet to be established. This study focused to identify novel polymorphisms in heat‐shock protein 70 (HSP70) gene and its possible association with post‐partum anoestrus in Murrah buffaloes. A 579‐bp fragment from 5′ untranslated region of HSP70 gene was amplified by polymerase chain reaction from blood genomic DNA of 614 animals maintained under similar management conditions. In phase‐I experiment, custom sequencing and restriction enzyme (RE) digestion of the amplified fragment were performed in 40 buffaloes with similar post‐partum oestrous conditions over previous consecutive three or more gestations—20 animals each showing post‐partum anoestrus (>120 days after parturition) and normal cyclicity (<65 days after parturition). While in phase‐II experiment, herd screening by RE analysis was performed in remaining 574 animals. Four transversions at T‐75G, C+31G, T+38G and C+97A and three transition mutations at T‐153C, T+33C and A+44G positions were observed. Polymorphism at T+38G site revealed significant (p < .05) variation, where homozygous G was present only in post‐partum anoestrous animals while nucleotide T was present randomly in both groups of phase‐I animals whereas phase‐II experiments revealed homozygous G in 55 animals. Regression analysis in relation to average post‐partum interval against genotypic frequencies at T+38G also depicted significant association. HSP70 gene polymorphism at T+38G position can therefore be used as genetic marker for excluding probable post‐partum anoestrous buffaloes from herd for breeding programmes.     

An imputation‐based genome‐wide association study on traits related to male reproduction in a White Duroc × Erhualian F2 population

Boar reproductive traits are economically important for the pig industry. Here we conducted a genome‐wide association study (GWAS) for 13 reproductive traits measured on 205 F₂ boars at day 300 using 60 K single nucleotide polymorphism (SNP) data imputed from a reference panel of 1200 pigs in a White Duroc × Erhualian F₂ intercross population. We identified 10 significant loci for seven traits on eight pig chromosomes (SSC). Two loci surpassed the genome‐wide significance level, including one for epididymal weight around 60.25 Mb on SSC7 and one for semen temperature around 43.69 Mb on SSC4. Four of the 10 significant loci that we identified were consistent with previously reported quantitative trait loci for boar reproduction traits. We highlighted several interesting candidate genes at these loci, including APN, TEP1, PARP2, SPINK1 and PDE1C. To evaluate the imputation accuracy, we further genotyped nine GWAS top SNPs using PCR restriction fragment length polymorphism or Sanger sequencing. We found an average of 91.44% of genotype concordance, 95.36% of allelic concordance and 0.85 of r² correlation between imputed and real genotype data. This indicates that our GWAS mapping results based on imputed SNP data are reliable, providing insights into the genetic basis of boar reproductive traits.     

In vitro anti‐tubulin effects of mebendazole and fenbendazole on canine glioma cells

Benzimidazole anthelmintics have reported anti‐neoplastic effects both in vitro and in vivo. The purpose of this study was to evaluate the in vitro chemosensitivity of three canine glioma cell lines to mebendazole and fenbendazole. The mean inhibitory concentration (IC₅₀) (±SD) obtained from performing the MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] assay after treating J3T, G06‐A, and SDT‐3G cells for 72 h with mebendazole were 0.030 ± 0.003, 0.080 ± 0.015 and 0.030 ± 0.006 μM respectively, while those for fenbendazole were 0.550 ± 0.015, 1.530 ± 0.159 and 0.690 ± 0.095 μM; treatment of primary canine fibroblasts for 72 h at IC₅₀ showed no significant effect. Immunofluorescence studies showed disruption of tubulin after treatment. Mebendazole and fenbendazole are cytotoxic in canine glioma cell lines in vitro and may be good candidates for treatment of canine gliomas. Further in vivo studies are required.     

An association study using imputed whole‐genome sequence data identifies novel significant loci for growth‐related traits in a Duroc × Erhualian F2 population

The average daily gain (ADG) and body weight (BW) are very important traits for breeding programs and for the meat production industry, which have attracted many researchers to delineate the genetic architecture behind these traits. In the present study, single‐ and multi‐trait genome‐wide association studies (GWAS) were performed between imputed whole‐genome sequence data and the traits of the ADG and BW at different stages in a large‐scale White Duroc × Erhualian F₂ population. A bioinformatics annotation analysis was used to assist in the identification of candidate genes that are associated with these traits. Five and seven genome‐wide significant quantitative trait loci (QTLs) were identified by single‐ and multi‐trait GWAS, respectively. Furthermore, more than 40 genome‐wide suggestive loci were detected. On the basis of the whole‐genome sequence association study and the bioinformatics analysis, NDUFAF6, TNS1 and HMGA1 stood out as the strongest candidate genes. The presented single‐ and multi‐trait GWAS analysis using imputed whole‐genome sequence data identified several novel QTLs for pig growth‐related traits. Integrating the GWAS with bioinformatics analysis can facilitate the more accurate identification of candidate genes. Higher imputation accuracy, time‐saving algorithms, improved models and comprehensive databases will accelerate the identification of causal genes or mutations, which will contribute to genomic selection and pig breeding in the future.     

Characterization of bovine neutrophil β2‐adrenergic receptor function

LaBranche, T. P., Ehrich, M. F., Eyre, P. Characterization of bovine neutrophil β₂‐adrenergic receptor function. J. vet. Pharmacol. Therap. doi: 10.1111/j.1365‐2885.2009.01143.x. This study compares bovine leukocyte β‐adrenergic receptor densities to that of the rat, demonstrates for the first time a functional β₂‐adrenergic receptor signaling pathway in steer neutrophils, and investigates the effect of an inflammatory stimulus on that signaling pathway. The β₁‐/β₂‐adrenergic antagonist ^[3H]CGP‐12177 demonstrated that rat lymphocyte specific binding‐site density was highest, followed by steer and dairy cow lymphocytes, and lastly steer and dairy cow neutrophils. The β₂‐adrenergic agonist terbutaline stimulated steer neutrophil adenosine 3,5‐cyclic monophosphate (cAMP) production, an effect increased by inclusion of ≥1 × 10^?8 m phorbol 12‐myristate 13‐acetate (PMA), an activator of protein kinase C. Both terbutaline and the nonselective phosphodiesterase inhibitor 3‐isobutyl‐1‐methylxanthine (IBMX) independently decreased steer neutrophil superoxide anion production in a concentration‐dependent manner, with 1 × 10^?4 m IBMX enhancing both the potency and efficacy of the terbutaline effect (up to 74% reduction in superoxide anion production). Superoxide anion production was also reduced by the synthetic cAMP analog 8‐bromo‐cAMP, which increased the potency of the IBMX effect on superoxide anion production. Taken together, these data demonstrate the presence of a β₂‐adrenergic receptor signaling pathway in bovine neutrophils much like that described in other animal species, as well as the potential for an inflammatory stimulus to alter its function.     

Tolerance evaluation of overdosed dietary levels of 25‐hydroxyvitamin D3 in growing piglets

Forty‐eight, cross‐bred (GL × LW × P) piglets were used in a 42‐day tolerance trial to assess the effects of feeding diets supplemented with vitamin D or increasing levels of 25‐hydroxyvitamin D₃ (25‐OH‐D₃). Six‐week‐old piglets (24 castrate males, 24 females) were used. Two replicate groups of 6 piglets were randomized by weight and allocated to four dietary treatments. The control group (T1) was supplemented with 50 μg vitamin D₃/kg feed. The experimental groups received 25‐OH‐D₃ at the recommended dose (T2: 50 μg/kg = 1x), at 250 μg/kg (T3: 5x) or at 500 μg/kg (T4: 10x) respectively. Feed intake and daily weight gain were measured weekly, and the animals were examined by a veterinarian daily. After 42 days, body mass, blood, urine, bone and tissue samples were analysed and a pathology examination conducted. Dietary treatments had no significant effect on final body mass or daily weight gain. The 25‐OH‐D₃ plasma concentration in T1 was 17 ± 3 ng/ml (mean ± SD) while the respective values of the experimental groups were significantly increased in T2, T3 and T4. Tissue concentrations of 25‐OH‐D₃ were higher in liver and muscle for T3 and T4 and in skin for T4 than in T1. However, neither gross pathology nor histology, nor blood and urine characteristics, nor bone parameters were affected by dietary treatments. Weight of organs as well as dry matter, ash and calcium content of kidneys remained unaffected by dietary 25‐OH‐D₃ intake. Furthermore, no changes were observed for general indicators of health. The results of this study demonstrated that feeding piglets with 25‐OH‐D₃ at 5 or 10 times the recommended level had no adverse effects on any of the biological parameters measured. It was concluded that 25‐OH‐D₃ can be regarded as a supplement with a very high safety margin when used at the recommended level.     

Expression analysis of an α‐1, 3‐galactosyltransferase,an enzyme that creates xenotransplantation‐related α–Gal epitope,in pig preimplantation embryos

α‐1,3‐Galactosyltransferase (α‐GalT), an enzyme creating Galα1‐3Gal (α‐Gal) epitope on the cell surface in some mammalian species such as pigs, is known to be a key factor that causes hyperacute rejection upon transplantation from pigs to humans. To establish the RNA interference‐based suppression of endogenous α‐GalT messenger RNA (mRNA) synthesis in porcine preimplantation embryos, we determined the suitable embryonic stage at which stage such approach is possible by using the semi‐quantitative RT‐PCR (qRT‐PCR) and the cytochemical method using a fluorescence‐labeled Bandeiraea simplicifolia Isolectin B₄ (BS‐I‐B₄). Staining with BS‐I‐B₄ demonstrated that α‐Gal epitope expression was first recognized at the 8‐cell stage, and increased up to the hatched blastocyst stage. Single embryo‐based qRT‐PCR also confirmed this pattern. These results indicate that creation of α‐Gal epitope is proceeded by de novo synthesis of α‐GalT mRNA in porcine preimplantation embryos with peaking at the blastocyst stage.     

Molecular analysis of the sex chromosomes of the platyfish Xiphophorus maculatus: Towards the identification of a new type of master sexual regulator in vertebrates

In contrast to mammals and birds, fish display an amazing diversity of genetic sex determination systems, with frequent changes during evolution possibly associated with the emergence of new sex chromosomes and sex‐determining genes. To better understand the molecular and evolutionary mechanisms driving this diversity, several fish models are studied in parallel. Besides the medaka (Oryzias latipes Temminck and Schlegel, 1846) for which the master sex‐determination gene has been identified, one of the most advanced models for studying sex determination is the Southern platyfish (Xiphophorus maculatus, Günther 1966). Xiphophorus maculatus belongs to the Poeciliids, a family of live‐bearing freshwater fish, including platyfish, swordtails and guppies that perfectly illustrates the diversity of genetic sex‐determination mechanisms observed in teleosts. For X. maculatus, bacterial artificial chromosome contigs covering the sex‐determination region of the X and Y sex chromosomes have been constructed. Initial molecular analysis demonstrated that the sex‐determination region is very unstable and frequently undergoes duplications, deletions, inversions and other rearrangements. Eleven gene candidates linked to the master sex‐determining gene have been identified, some of them corresponding to pseudogenes. All putative genes are present on both the X and the Y chromosomes, suggesting a poor degree of differentiation and a young evolutionary age for platyfish sex chromosomes. When compared with other fish and tetrapod genomes, syntenies were detected only with autosomes. This observation supports an independent origin of sex chromosomes, not only in different vertebrate lineages but also between different fish species.     

EVALUATION OF QUANTITATIVE THYROID SCINTIGRAPHY FOR DIAGNOSIS AND STAGING OF DISEASE SEVERITY IN CATS WITH HYPERTHYROIDISM: COMPARISON OF THE PERCENT THYROIDAL UPTAKE OF PERTECHNETATE TO THYROID‐TO‐SALIVARY RATIO AND THYROID‐TO‐BACKGROUND RATIOS

Thyroid scintigraphy is commonly used for evaluation of cats with hyperthyroidism, with the thyroid‐to‐salivary ratio (T/S) being the most common method to quantify the degree of thyroid activity and disease. Calculation of thyroid‐to‐background ratios (T/B) or percent thyroidal uptake of ^99mTcO^?₄ (TcTU) has only been reported in a few studies. The purpose of this prospective, cross‐sectional study was to evaluate a number of quantitative scintigraphic indices as diagnostic tests for hyperthyroidism, including the T/S, three different T/B, TcTU, and estimated thyroid volume. Of 524 cats referred to our clinic for evaluation of suspected hyperthyroidism, the diagnosis was confirmed (n = 504) or excluded (n = 20) based on results of a serum thyroid panel consisting of thyroxine (T₄), triiodothyronine (T₃), free T₄ (fT₄), and thyroid‐stimulating hormone (TSH) concentrations. In the hyperthyroid cats, median values for TcTU, T/S, and three T/B ratios were all significantly higher (P < 0.001) than values in euthyroid suspect cats or clinically normal cats. All scintigraphic parameters were relatively sensitive and specific as diagnostic tests for hyperthyroidism, but the T/S ratio had the highest test accuracy. The T/S ratio correlated strongly with the TcTU (r = 0.85). However, the TcTU had a higher and more significant correlation (P < 0.01) with serum T₄ (r = 0.76 vs. 0.64), T₃ (r = 0.77 vs. 0.64), and estimated thyroid volume (r = 0.62 vs. 0.38). Overall, calculation of TcTU is an accurate diagnostic test, but also appears to be the best parameter to predict the functional volume and metabolic activity of the feline adenomatous thyroid gland.     

Embryonic and post‐embryonic responses to high‐elevation hypoxia in a low‐elevation lizard

Low‐elevation species can migrate toward higher elevations to survive in a warming world. However, animals’ responses to hypoxia when migrating to high elevations have rarely been addressed. To identify the response of low‐elevation lizards to high‐elevation hypoxia, we collected field body temperatures (T_fb) and operative temperatures (T_e) of lizards (Eremias argus) from a low‐elevation population (1036 m) and a high‐elevation population (2036 m), and then determined adult thermal physiology, embryonic development, and hatchling phenotypes after acclimating low‐elevation lizards and incubating their eggs in conditions mimicking the low‐elevation oxygen condition (18.5% O₂) and high‐elevation oxygen (hypoxic) condition (16.5% O₂). Our study revealed that T_fb and T_e were higher for the low‐elevation population compared to the high‐elevation population. We also found adults from low elevation acclimated to hypoxia preferred lower body temperatures, but did not show changes in locomotor performance or growth. In addition, hypoxia did not affect embryonic development (hatching time and success) or hatchling phenotypes (body size and locomotor performance). These results suggest that adult lizards from low elevations can respond to hypoxia‐induced stress when migrating to high elevations by behaviorally thermoregulating to lower body temperatures in order to sustain normal functions. Similarly, low‐elevation embryos can develop normally (with unchanged hatching success and offspring phenotypes) under the high‐elevation hypoxic condition. This study highlights that low‐elevation populations of a species that inhabits a range of elevations can buffer the impact of high‐elevation hypoxic conditions to some degree and thus attain similar fitness to the source population.     

A biochemical and immunological comparative study on Trypanosoma equiperdum and Trypanosoma evansi

Trypanosoma equiperdum and Trypanosoma evansi were purified by three or four cycles of low-speed centrifugation and final filtration through DEAE cellulose. The purified trypanosomes were used in comparative biochemical and immunological studies. Comparative polypeptide pattern analysis revealed that T. equiperdum showed 21 polypeptide bands, whose M _r ranged from >200 to 14.8 kDa. T. evansi showed 25 polypeptide bands in the M _r range 97–14.8 kDa. The main differences were associated with the presence of secondary bands, relative intensity and the number of bands. Both species gave seven glycoprotein bands; those of 97 and 68 kDa were present in T. equiperdum but absent in T. evansi. Bands of 61 and 28 kDa were present in T. evansi but not in T. equiperdum. Anti-T. equiperdum sera recognized four homologous antigens and cross-reacted with three antigens of T. evansi. Anti-T. evansi sera recognized three homologous antigens and cross-reacted with four T. equiperdum antigens. Four identical proteolytic protease bands were present for both species, while only one surface protein was detected for each species: 66 kDa for T. equiperdum and 62 kDa for T. evansi.     

Non‐gelatinized starch influences the deposition of n‐3 fatty acids in the muscle of a tropical freshwater fish,Labeo rohita

A 60‐day feeding trial was conducted to study the influence of gelatinized (G) to non‐gelatinized (NG) starch ratio in the diet on fatty acids profiles and oxidative status in Labeo rohita fingerlings. Two hundred and thirty‐four fingerlings (average weight: 2.53 g) were distributed in six treatment groups with each of three replicates. Six semi‐purified diets either containing NG and/or G corn starch (42.43%) viz., T₁ (100% NG and 0% G starch), T₂ (80% NG and 20% G starch), T₃ (60% NG and 40% G starch), T₄ (40% NG and 60% G starch), T₅ (20% NG and 80% G starch) and T₆ (0% NG and 100% G starch) was fed to respective groups. Catalase, superoxide dismutase and malic enzyme activities decreased linearly with the increasing level of G starch, whereas reverse trend was found for glucose‐6‐phosphate dehydrogenase. Total saturated fatty acids in muscle increased with the increasing level of G starch in the diet. Total n‐3 fatty acids decreased linearly with the increasing level of G starch in the diet. Among the n‐3 fatty acids, linolenic acids content was more in NG starch fed group. Similarly, eicosapentaenoic acid contents gradually decreased with increasing level of G starch content. The n‐6/n‐3 ratio was higher in G starch fed group. This suggests that dietary starch type may be manipulated for quality improvement of fish flesh.     

Effect of non‐starch‐polysaccharide‐degrading enzymes as feed additive on the rumen bacterial population in non‐lactating cows quantified by real‐time PCR

The effects of non‐starch‐polysaccharide‐degrading enzymes, added to a maize silage‐ and grass silage‐based total mixed ration (TMR) at least 14 h before feeding, on the rumen bacterial population were investigated. Six non‐lactating Holstein Friesian cows were allocated to three treatment groups using a duplicate 3 × 3 Latin square design with three 31‐day periods (29 days of adaptation and 2 days of sampling). Treatments were control TMR [69% forage and 31% concentrates on a dry matter (DM) basis] or TMR with 13.8 or 27.7 ml/kg of feed DM of Roxazyme G2 liquid with activities (U/ml enzyme preparation) of xylanase 260 000, β‐glucanase 180 000 and cellulase 8000 (DSM Nutritional Products, Basel, Switzerland). The concentrations of 16S rDNA of Anaerovibrio lipolytica, Fibrobacter succinogenes, Prevotella ruminicola, Ruminococcus flavefaciens, Selenomonas ruminantium and Treponema bryantii, and their relative percentage of total bacteria in rumen samples obtained before feeding and 3 and 7 h after feeding and from two rumen fractions were determined using real‐time PCR. Sampling time had only little influence, but bacterial numbers and the composition of the population differed between the transition layer between rumen fluid and the fibre mat (fraction A) and the rumen fluid (fraction B) highlighting the importance to standardize sampling. The 16S rDNA copies of total bacteria and the six bacterial species as well as the population composition were mainly unaffected by the high levels of exogenous enzymes supplemented at all sampling times and in both rumen fractions. Occasionally, the percentages of the non‐fibrolytic species P. ruminicola and A. lipolytica changed in response to enzyme supplementation. Some increases in the potential degradability of the diet and decreases in lag time which occurred collaterally indicate that other factors than changes in numbers of non‐particle‐associated bacteria are mainly responsible for the effects of exogenous enzymes.