Diagnosis of caseous lymphadenitis by ELISA in naturally infected goats from Venezuela

Due to the absence of previous reports, the goal of this work was to detect caseous lymphadenitis (CLA) in goat flocks from Venezuela using an indirect immunoenzymatic assay (ELISA). Eighteen farms were randomly selected in Falcon State, North-Western Venezuela. Blood samples were taken from 259 goats, 65 of them with abscesses. Experimental inoculations were made to healthy kids with 0.5 mL inocula containing 4.7 × 10⁵ of Corynebacterium pseudotuberculosis to observe the kinetics of antibody response. Immunoenzymatic assays were carried out using exotoxin of C. pseudotuberculosis as antigen. Antibody response in experimentally inoculated animals was detected 2 weeks after infection. Of 259 field goat sera, 55.98% were positive by ELISA. Of 65 goats with abscesses, 67.69% had CLA demonstrated by bacteriological methods; from these, 72.73% showed antibodies by ELISA. Of the remaining goats negative to CLA, 47.62% had antibodies by ELISA. Sensitivity was calculated in 72.73% and specificity in 67.74%. The immunoenzymatic assay applied in this research could be useful to detect CLA in naturally infected goat flocks from Venezuela.     

A primary Corynebacterium pseudotuberculosis low dose infection in alpacas (Lama pacos) protects against a lethal challenge exposure

Corynebacterium pseudotuberculosis is the agent of alpaca's lymphadenitis. The present study was to demonstrate the effect of a primary infection with low (1.1 × 10³), moderate (1 × 10⁴), and high (1.2 × 10⁵) doses of C. pseudotuberculosis against a significant higher challenge dose of 9 × 10⁸ CFU of C. pseudotuberculosis. Three groups of 4 healthy male alpacas were inoculated subcutaneously (SC) in the left flank behind the costal arch with the above doses of bacteria. A fourth group of 4 alpacas was sham inoculated with phosphate buffered saline as control. After 5 weeks all animals were challenged with a dose of 9 × 10⁸ CFU of C. pseudotuberculosis inoculated SC in the right flank. The alpacas were clinically inspected for local and regional abscesses, body temperature and behavior changes. The primary infected alpacas had a febrile response, and abscesses at the inoculation point and regional lymph nodes. However, after challenge, the primary infected animals showed no superficial lesions or febrile response. In contrast, the immune naïve alpacas from group D developed a severe disease characterized by fever, abscesses in regional lymphnodes, and in one alpaca a subcutaneous edema and sudden death 2 weeks after exposure. In addition, primary infected alpacas had a robust antibody response against C. pseudotuberculosis cell wall antigen with significant differences with respect the naïve challenged alpacas. At necropsy, the primary infected alpacas had abscesses only in the regional or internal renal-lymph nodes from the left or primary inoculation side of the body, with no lesions in the right challenged side. In contrast, the primary sham inoculated alpacas had abscesses in the regional and internal lymph nodes from the right challenged side. This work showed that a primary infection with at least 1.1 × 10³ viable C. pseudotuberculosis induces protection against a second high dose exposure to this bacterium. These results will be useful for further study of prevention methods to control lymphadenitis in alpacas.     

Immunological characterization of diphtheria toxin recovered from Corynebacterium pseudotuberculosis

Diphtheria toxin (DT) is a potent toxin produced by the so-called diphtheria group which includes Corynebacterium diphtheriae (C. diphtheriae), Corynebacterium ulcerans (C. ulcerans), and Corynebacterium pseudotuberculosis (C. pseudotuberculosis). The present investigation is aimed to study in detail the production of DT by C. pseudotuberculosis. Twenty isolates were obtained from sheep diseased with caseous lymphadenitis (CLA) and twenty-six isolates were obtained from 26 buffaloes diseased with oedematous skin disease (OSD). All isolates were identified by standard microbiological and DT production was assayed serologically by modified Elek test and immunoblotting. All sheep isolates were nitrate negative, failed to hydrolyze starch and could not produce DT, while all buffalo isolates (biotype II) revealed positive results and a specific band of 62 kDa, specific to DT, was resulted in all concentrated cell fractions (CF), but was absent from non-toxigenic biotype I isolates. At the same time, another band of 31 kDa specific to the PLD gene was obtained with all isolates of biotype I and II. Moreover, all isolates showed positive synergistic hemolytic activity and antagonistic hemolysis with β-hemolytic Staphylococci. The obtained results also indicated that C. pseudotuberculosis could be classified into two strains; non-toxigenic biotype I strain, which failed to produce DT as well as being negative to nitrate and starch hydrolysis, and toxigenic biotype II strain, which can reduce nitrate, hydrolyze starch as well as produce DT.     

Trace mineral status and liver and blood parameters in sheep without mineral supply compared to local roe deer (Capreolus capreolus) populations

General health, clinical-chemical blood analysis and copper (Cu), zinc (Zn), selenium (Se) and vitamin E concentrations in plasma and liver tissue (wet weight, ww) of two extensive grazing sheep flocks without mineral supply were compared to the status of local roe deer (Capreolus capreolus) populations (liver samples). Both sheep flocks were classified as healthy except for a remarkable variation in body weight and a slight foot rot infection in one flock. Hematology of sheep was normal, and total protein and creatinine as well as activities of creatin kinase, aspartat-amino-transferase, alkaline phosphatase and gamma-glutamyl-transferase in plasma were within reference levels. The mean of glutamate dehydrogenase (13.8 U/l) was slightly elevated in one flock. Mean liver concentrations of Zn (38.9 and 43.5 mg/kg ww) and Cu (111 and 87.5 mg/kg ww) in sheep flocks were higher compared to the respective roe deer populations (27.5 and 36.3 mg Zn/kg ww; 18.3 and 28.6 mg Cu/kg ww). This is supposed to be caused by differences in Cu and Zn metabolism in sheep and roe deer. Selenium deficiency was diagnosed in liver samples of both sheep flocks (0.21 and 0.23 mg/kg ww). There were neither significant differences compared to roe deer (0.21 and 0.27 mg Se/kg ww) nor differences depending on location. Correlations between plasma and liver concentrations of Cu, Zn and Se were not significant in sheep. Means of vitamin E in liver samples (30.6 and 41.8 mg/kg ww) were higher in roe deer populations. This may be caused by the opportunity of selective browsing for wild ruminants, which allows access to younger plants which are higher in vitamin E.     

Gender discrimination in undernutrition with mediating factors among Bengalee school children from Eastern India

This study was undertaken to determine age and sex variations in the prevalence of underweight and stunting, and to assess the impact of some socio-economic variables on undernutrition among 6–16 year old school children of Bengalee ethnicity in Chapra, West Bengal, India. The subjects were selected randomly from various schools and madrassas of the Chapra Block. A total of 725 children (342 boys and 383 girls) aged 6–16 years were measured and data on their socio-economic status were collected. Age and sex combined rates of underweight and stunting were 44.40% and 37.20%, respectively. Weight-for-age Z-score (WAZ) showed significant association with per-capita income (PCI) among boys (F = 5.45) and girls (F = 8.14). Height-for-age Z-score (HAZ) has also shown the association with per-capita income among boys (F = 4.43) and girls (F = 9.69). The WAZ was significantly associated with fathers’ educational status (FOS) (t = ?2.95) and the number of living rooms (NLR) (t = ?2.91) among girls. The HAZ showed significant association with number of siblings (NS) among girls (F = 4.25). Linear regression analyses revealed that NLR (t = 2.04) and NS (t = 1.95) had a significant impact on HAZ among boys. Among girls, PCI (t = 3.38), FOS (t = 2.87) and NLR (t = 2.81) had a significant impact on WAZ and also PCI (t = 3.28) and FOS (t = 2.90) had a significant impact on HAZ. NLR had significant associations with underweight (χ² = 3.59) and stunting (χ² = 4.20) among boys. Among girls, PCI had significant associations with underweight (χ² = 11.15) and stunting (χ² = 11.64). FOS also showed significant associations with underweight (χ² = 8.10) as well as stunting (χ² = 8.28) among girls. NLR showed a significant association with underweight (χ² = 7.75). Logistics regression analyses revealed that FOS (Wald = 8.00) and NLR (Wald = 4.09) were significant predictors of stunting among boys. Among girls, PCI was a significant predictor of underweight (Wald = 10.95) as well as stunting (Wald = 10.45). FOS, NLR and NS were also significant predictors of stunting (Wald = 8.16), underweight (Wald = 7.68) and stunting (Wald = 6.97) respectively. The present study revealed that the nutritional status of the children was unsatisfactory and it is of paramount importance not only to increase the amount of food supplementation given but also to promote gender equality.     

Determination of prediction equations for estimating body weight of Zulu (Nguni) sheep

Data on linear body measurements (LBM) of 403 sheep collected in three areas of KwaZulu-Natal were utilized to develop a prediction equation for live body weight of Zulu sheep. Data were collected on live weight (LW), heart girth (HG), wither height (WH) and scrotum circumference (SC) on sheep of all ages. The age of sheep was estimated by dentition. The analysis of variance showed that age and sex were important factors contributing to variation in LW of Zulu sheep. Phenotypic correlation coefficients and regression equations of LW on HG, WH and SC were computed within different age groups (milk set of teeth, one pair, two pairs and the three and four pairs of incisors). Low correlation coefficients (r = 0.21–0.48) between LW, HG and WH were found among the pregnant ewes. The relationship between LBM and LW was stronger (r = 0.66–0.86) for males than among females (r = 0.42–0.75). The cubic polynomial of HG was the best fit (R² = 0.76) for the live weight prediction of young sheep with milk set of teeth. The combination of HG and WH produced the best fit for the two tooth and above males and non-pregnant females. The LW prediction equations for pregnant females were not reliable (R² = 0.05–0.26). The SC was more precise (R² = 0.61–0.80) when estimating the live weight of young males (<15–22-month-old) than of the older rams (R² = 0.23–0.56). It was concluded that LW of Zulu sheep can be reasonably estimated using the HG and WH. A table could be constructed for the farmers to estimate the LW of their animals.     

Persistent efficacy of long-acting moxidectin for control of trichostrongylid infections of sheep

The objective was to evaluate long-acting moxidectin (Cydectin^® 2% LA for sheep) against trichostrongylids in sheep. We performed a blocked, parallel, controlled clinical trial. We included 45 ewe-lambs, allocated into treated (n = 30, 1.0 mg moxidectin per kg bw, subcutaneously at base of ear on D 0) or controls (n = 15). Animals had been naturally infected (pre-treatment geometric mean epg counts: 233 and 249, respectively) and throughout the study grazed at the same pasture. Fortnightly, we collected faecal samples and performed established parasitological techniques (epg counts, coprocultures). We used analysis of covariance for post-treatment results, using pre-treatment counts as covariate, treatment as fixed effect, block as random effect. Geometric mean epg counts in treated animals were 0 up to D 112, 1 on D 126, 4 on D 140 and 13 on D 161. Respective figures for controls were 369–820 up to D 140 and 168 on D 161 (p < 0.05). Persistent efficacy (>90%) was 140 d for Teladorsagia, 119 d for Haemonchus and 115 d for Trichostrongylus. This is the first reported clinical trial of efficacy of long-acting moxidectin in sheep in Europe. Cydectin 2% LA was found to be effective against trichostrongylids of sheep. The particularly long persistent efficacy can provide new possibilities in formulating anthelmintic programs for sheep.     

The relationship between the volatile fatty acids supply and the nitrogen retention in growing sheep nourished by total intragastric infusions

Twelve four-month old Suffolk × Small-tail-Han male sheep (live weight 21–26 kg), fitted with rumen and abomasum fistulas and nourished by total intragastric infusions, were used to study the relationship between the volatile fatty acids (VFA) supply and the nitrogen (N) retention in sheep. The animals were randomly divided into four groups and four levels of mixed VFA energy (the molar proportion of acetic acid, propionic acid and butyric acid was 65:25:10), i.e. 333, 378, 423 and 468 kJ kg^?1 W^0.75 d^?1, were infused into the rumen, as treatments I, II, III and IV, respectively. The results showed that the N retention was significantly increased (P < 0.05) with the VFA infusion level. Significant regression relationship was found between the VFA supply (x, g d^?1) and the N retention (y, mg d^?1): y = 2.75x ? 403, r² = 0.86, n = 12, P < 0.01. It was concluded that to get efficient utilization of dietary N and high N retention in sheep, it is necessary to supply enough dietary energy.     

Tumor Necrosis Factor-α, Interleukins-12(p40), 6, and 10 levels in cerebrospinal fluid and outcome prediction in Ossimi sheep with encephalitic listeriosis

Encephalitic listeriosis in sheep is a life-threatening disease. However, little is known about the cytokine response and their predictive value in this disease. The aim of present study was to assess the prognostic significance of Tumor Necrosis Factor-α (TNF-α), Interleukin-12(p40) (IL-12 p40), Interleukin-6 (IL-6), and Interleukin 10 (IL-10) levels in cerebrospinal fluid (CSF) in sheep with encephalitic listeriosis. Fifty-nine ewes in 14 flocks were diagnosed clinically as having listeriosis. CSF was collected and subjected to bacteriological examination and estimation of selected cytokines. Twenty-eight ewes were confirmed to be infected with Listeria monocytogenes. Based on antimicrobial sensitivity test, sheep were treated and the outcome was recorded as survivors (n = 10) and non-survivors (n = 18). Cutoff points for CSF cytokines were determined by Receiver operating characteristic analysis (ROC). Association between levels of CSF cytokines and outcome of listeriosis was assessed by logistic regression. TNF-α, IL-6 and IL-12(p40) levels as well as TNF-α/IL-10 ratio were significantly higher in non-survivors than survivors (p = 0.002, 0.0021, 0.0033, and 0.001, respectively). However, IL-10 level was significantly lower in non-survivors than survivors (p = 0.0058). ROC analysis revealed that IL-6 and TNF-α/IL-10 ratio had the highest AUC values (0.98, 0.984, respectively). Final multivariate logistic regression model showed that TNF-α/IL-10 ratio was the only variable that has predictive value for mortality in diseased sheep (p: 0.001; OR: 7.2; 95% CI: 5.7–9.8). TNF-α showed a positive correlation with IL-12β (r = 0.917) and IL-6 (r = 0.965). IL-12 (p40) showed also a positive correlation with IL-6 (r = 0.906). However, IL-10 showed a negative correlation with TNF-α (r = −0.915), IL-12(p40) (r = −0.790), and IL-6 (r = −0.902). In conclusion, TNF-α/IL-10 ratio may provide predictive information about outcome of encephalitic listeriosis in sheep.     

A missense mutation (c.184C > T) in ovine CLN6 causes neuronal ceroid lipofuscinosis in Merino sheep whereas affected South Hampshire sheep have reduced levels of CLN6 mRNA

The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are a group of fatal recessively inherited neurodegenerative diseases of humans and animals characterised by common clinical signs and pathology. These include blindness, ataxia, dementia, behavioural changes, seizures, brain and retinal atrophy and accumulation of fluorescent lysosome derived organelles in most cells. A number of different variants have been suggested and seven different causative genes identified in humans (CLN1, CLN2, CLN3, CLN5, CLN6, CLN8 and CTSD). Animal models have played a central role in the investigation of this group of diseases and are extremely valuable for developing a better understanding of the disease mechanisms and possible therapeutic approaches. Ovine models include flocks of affected New Zealand South Hampshires and Borderdales and Australian Merinos. The ovine CLN6 gene has been sequenced in a representative selection of these sheep. These investigations unveiled the mutation responsible for the disease in Merino sheep (c.184C > T; p.Arg62Cys) and three common ovine allelic variants (c.56A > G, c.822G > A and c.933_934insCT). Linkage analysis established that CLN6 is the gene most likely to cause NCL in affected South Hampshire sheep, which do not have the c.184C > T mutation but show reduced expression of CLN6 mRNA in a range of tissues as determined by real-time PCR. Lack of linkage precludes CLN6 as a candidate for NCL in Borderdale sheep.     

Screening for Booroola (FecB) and Galway (FecXG) mutations in Indian sheep

This study reports the status of the Booroola (FecB) and Galway (FecX^G) mutations in Indian sheep breeds. The Kendrapada sheep (n = 46) was genotyped for the presence of FecB and FecX^G mutations, while the Garole (n = 34), Malpura (n = 30), and Decanni sheep (n = 15) for the FecX^G mutation. The FecB and FecX^G genotyping was carried out by forced restriction fragment length polymorphism PCR technique. In the present study, FecB mutation was discovered in the Kendrapada sheep of Orissa, which is now the second prolific sheep of India after the Garole. Out of 46 individuals of Kendrapada sheep, 26 were homozygous (BB), 15 heterozygous (B+) and 5 non-carriers (++) for the FecB mutation. The frequency of the FecB allele in this sample was about 0.73. Results indicated that the frequency of the FecB mutation is high, but the gene is not fixed in the population as reported in Garole sheep. None of sheep breeds carried the FecX^G mutation. The discovery of the FecB mutation in Kendrapada sheep will facilitate the use of FecB allele in improving the prolificacy of non-prolific sheep breeds of India.     

In vivo and in situ digestibilities and nitrogen balance of Buddleia skutchii as a sole component and mixed with Pennisetum clandestinum in sheep diets

The objective of this work was to evaluate the intake (I) of all Buddleia skutchii foliage (BSF), collected from three different sites (places: A, B and C) in feeding sheep (phase 1), and in combination with Pennisetum clandestinum (Pc) (phase 2). Trials of feed intake, in vivo dry matter digestibility (IVDMD), and digestibilities of crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and nitrogen balance were carried out during phase 1. Diet was given to nine Creole male lambs of 23(±3) kg body weight (BW), located in individual metabolism crates. During phase 2, four treatments were evaluated (T1 = 100% Pc, T2 = 20% BSF + 80% Pc, T3 = 40% BSF + 60% Pc and T4 = 60% BSF + 40% Pc) in a 4 × 4 Latin square design, using four ruminal cannulated Creole male rams of 50 ± 2 kg BW and remained in individual metabolism crates. In vivo digestibility (DM, CP and gross energy (GE)) and kinetics of in situ disappearance of DM were evaluated. The results in phase 1 indicated that BSF from Site B had relatively higher CP (14.9%), ADF (50.6%) content and nitrogen retention (1.93 g d⁻¹). On the other hand, the lambs showed intake levels of 376.29 g d⁻¹ DM, 56.21 g d⁻¹ CP, 206.24 g d⁻¹ ADF and 1418.21 kcal gross energy (P > 0.05); and digestibilities of 54.23% DM, 47.17% CP and 39.31% NDF. Therefore, BSF from Site B was selected for the second phase of this work as the best result. We observed that the nutrition quality of the diets was higher as a result of increasing BSF concentration in the diet. When sheep were fed on 40% BSF–60% Pc, DM intake levels of 900.6 g d⁻¹, and DM, CP and GE digestibility of 70.0, 52.82 and 55.1%, respectively, were observed. There were not significant differences among diets, and in situ disappearance, K_d and K_p, but the diet with 60% of BSF presented higher ruminal digestibility (42.3%). The results allowed to validate the local knowledge on BSF as a promising forage species, and showed that BSF should be incorporated in a sheep production system.     

Seroprevalence and risk factors associated with zoonotic parasitic infections in small ruminants in the Greek temperate environment

A cross-sectional serological study was carried out to screen the sheep and goat population of Thessaly, Greece for evidence of infection with Toxoplasma, Toxocara, Leishmania, and Echinococcus and to determine the risk factors related to herd characteristics, herd management practices, farmer status, and the bioclimatic variables associated with these zoonotic parasitic infections. A total of 540 sheep and goat serum samples were examined. The seroprevalence of infection in all examined animals was 24.5% for Toxoplasma, 32% for Toxocara, 0% for Leishmania and 85.9% for Echinococcus. The final logistic regression model showed that the species of small ruminant, herd size, anthelmintic treatment, class of anthelmintic treatment, grazing with other herds, educational level of farmer, elevation of farm location, and generalized land cover were associated with Toxoplasma gondii infections, while the species of small ruminant, farm type, anthelmintic treatment, class of anthelmintic treatment, rotation of grazing, age of farmer, elevation of farm location, and generalized land cover were associated with Toxocara canis infections. Antibodies to T. gondii were detected in 102 (28.3%) of 360 sheep and in 30 (16.8%) of 179 goats. Animals in small flocks (150–300 animals) had an approximately 0.42-fold lower risk of having positive cases of T. gondii among animals compared with large flocks (> 300 animals). Antibodies to T. canis were found in 155 (42.9%) of 361 sheep and 18 (10.1%) of 179 goats. The later finding constitutes the first report of seropositive goats to Toxocara. The risk of positivity for T. canis was 7.71-fold higher in sheep than in goats. Geographically, animals from plain areas had 2.9 and 2.01-fold higher risk of having positive cases of T. gondii and T. canis respectively. The significant bioclimatic variables (p < 0.05) associated with the occurrence locations of T. gondii infection were related to higher temperature, lower precipitation, and lower elevation compared to the absence locations of T. gondii. The significant bioclimatic variables (p < 0.05) associated with occurrence locations of T. canis infection were related to lower temperature and higher precipitation compared to absence locations of T. canis. These findings are useful to formulate appropriate control strategies for zoonotic parasites of sheep and goats in Greece and other areas with similar climatic conditions.     

A study on ectoparasites of sheep and goats in eastern part of Amhara region,northeast Ethiopia

A study on ectoparasites of small ruminants was carried out in three districts (woredas) of the eastern part of Amhara Regional State, Ethiopia, from November 2003 to March 2004, with the objectives of determining the prevalence of ectoparasites and identifying the potential risk factors associated with the problem. Out of 752 sheep and 752 goats examined, 50.5% of sheep and 56.4% of goats were infested with one or more ectoparasites. The ectoparasites identified in sheep were Damalina ovis (38.5%), Melophagus ovinus (12.5%), tick infestations (3.4%) and Linognathus spp. (2.4%). In goats, parasites such as Linognathus spp. (28.3%), ticks (22.2%), sarcoptic mites (6.1%) and Ctenocephalides spp. (8.1%) were identified. The prevalence of M. ovinus and D. ovis infestations in sheep were significantly (p < 0.05) higher in the highlands than both lowlands and midlands, which were almost similar. In goats, the risk of sarcoptic mange infestation in lowland and midland was 4.6 and 5.0 times higher than the highlands, respectively. The prevalence of Linognathus spp. in goats was significantly (p < 0.05) higher in the highlands than the lowlands. Both in sheep and goats, no significant difference between young and adult age groups was observed for any ectoparasites except for D. ovis in sheep in which adults (42.2%) were more affected than young animals (29.9%). The goats with poor body condition were 4.3 times at risk for sarcoptic mange (OR = 4.3, p < 0.05), Linognathus spp. (OR = 2.1, p < 0.05) and tick (OR = 1.6, p < 0.05) infestation than goats of good body condition. On the contrary, no significant variation (p > 0.05) in ectoparasite infestation of sheep was observed in relation to body condition. Favorable climates, poor management, poor awareness of farmers and poor animal health extension services are believed to have contributed for the widespread occurrence of ectoparasites.     

Holocene environmental changes as recorded by mineral magnetism of sediments from Anguli-nuur Lake,southeastern Inner Mongolia Plateau,China

Two cores, one 1141-cm long (An-S) and the other 885-cm long (An-A), were retrieved from Anguli-nuur Lake (41°18′–24′N, 114°20′–27′E, ～ 1315 masl), one of the largest lakes in the transition zone between a semi-humid and semi-arid climate parallel to the present limit of the southeast monsoon along the southeastern Inner Mongolia Plateau in north China. Mineral-magnetic parameters (χ_lf, ARM, IRM_300mT, SIRM and IRM_? 300mT) were measured on An-S and two additional parameters (χ_ARM and HIRM) and four inter-parametric ratios (χ_ARM/SIRM, IRM_300mT/SIRM, IRM_? 300mT/SIRM and SIRM/χ_lf) were calculated. Potential sources of these lake sediments (catchment soils and dune materials close to the lake and in a distant sand plain) were sampled, and the magnetic properties of the surface-material specimens were measured. A chronological model was developed for An-S by comparing and combining AMS¹⁴C dates of An-S with ¹³⁷Cs, ²¹⁰Pb and AMS¹⁴C dates of An-A. With the help of surface-material magnetism, the magnetic data of An-S in combination with particle size, TOC and C/N and pollen analyses indicate the environmental changes during the last ～ 10,000 years around this lake. Conditions began to ameliorate at 10,900 cal. yr BP (9600 ¹⁴C yr BP) and thus relatively wet and warm environments prevailed during 10,900–8900 cal. yr BP (9600–8000 ¹⁴C yr BP). The Holocene optimum or the wettest and warmest conditions, was during 8900–7400 cal. yr BP (8000–6500 ¹⁴C yr BP). The environment began to deteriorate from 7400 cal. yr BP (6500 ¹⁴C yr BP) and the driest and coolest conditions occurred during 2200–480 cal. yr BP. There may have been a minor amelioration after 480 cal. yr BP. The inferred changes in palaeoenvironmental conditions around Anguli-nuur Lake are broadly in agreement with those around most other sites on the Inner Mongolia Plateau.     

Incorporating sheep into dryland grain production systems: I. Impact on over-wintering larva populations of wheat stem sawfly,Cephus cinctus Norton (Hymenoptera: Cephidae)

Wheat stem sawfly (WSS), Cephus cinctus Norton (Hymenoptera: Cephidae) is the most damaging insect pest to Montana's $ 1 billion dollar per year grain industry. Current WSS control methods are either expensive, reduce wheat yields, or are not effective. Our objective was to compare burning, grazing, tilling, trampling and clipping wheat stubble fields on over-wintering WSS larval populations. Treatments were evaluated in three experiments using a randomized complete block design and four replications at each site. Eight, six, and two sites were used for Experiments 1, 2, and 3, respectively. Contrast statements were used to make pre-planned comparisons among treatments. For Experiment 1, treatments were fall tilled, fall grazed, spring grazed, fall and spring combined (Fall/Spr) grazed, and an untreated control. Five mature ewes were confined with electric fence to 111 m² plot for 24 h in the fall and spring grazed treatments resulting in a stocking rate of 452 sheep d/ha. For Fall/Spr, the stocking rate was 904 sheep d/ha. For Experiment 2, treatments were fall grazed, fall burned, fall tilled, and an untreated control. In Experiment 3, treatments were fall trampled, spring trampled, Fall/Spr trampled, hand clipped to a stubble height of 4.5 cm, and an untreated control. Trampled treatments were done at the same stocking rates as grazing treatments but sheep were muzzled to prevent intake. Wheat stem sawfly larval numbers were collected in the fall and spring, pre- and post-treatment, respectively, by collecting all plant material from three, 0.46 m lengths of row and counting the number of live larvae present. In Experiment 1, WSS mortality was greater (P < 0.01) for the mean of all grazed treatments (68.4%) than either control (43%) or tilled (47%) plots. Mortality did not differ (P = 0.75) between fall (67%) and spring (64%) grazed plots but was greater (P = 0.02) for Fall/Spr (74%). In Experiment 2, larva mortality was greater (P < 0.01) for fall grazed (63%) than burned plots (52%). In Experiment 3, WSS mortality was greater (P < 0.01) for the mean of all trampling treatments (57%) than either control (33%) or clipped (32%) plots. Mortality did not differ (P > 0.25) between fall (54%) and spring trampling (47%) but was greater (P = 0.01) for Fall/Spr (70.6%). No differences (P > 0.25) were detected for WSS mortality when grazing was compared to trampling. These results indicate the potential for using grazing sheep to control wheat stem sawfly infestations in cereal grain production systems.     

Bioconversion of post harvest crop residues and cattle shed manure into value-added products using earthworm Eudrilus eugeniae Kinberg

The post-harvest residues of some local crops, e.g. wheat (Triticum aestivum), millets (Penniseum typhoides and Sorghum vulgare), and a pulse (Vigna radiata) were subjected to recycle through vermicomposting by using the epigeic earthworm Eudrilus eugeniae Kinberg, under laboratory conditions. The crop residues were amended with animal dung; and three types of vermibeds were prepared: (i) millet straw (S. vulgare + Pennisenum typhoides in equal quantity) + sheep manure (1:2 ratio) (MS), (ii) pulse bran (Vigna radiata) + wheat straw (Triticum aestivum) + cow dung (1:1:2 ratio) (PWC), and (iii) mixed crop residues (mixing of all types crop residues, used in this study) + cow dung in 1:1 ratio (MCR + CD). The fourth treatment was cattle shed manure (CSM). At the end, ready vermicompost showed lower organic C content and higher concentrations of other important plant nutrients. Organic C content decreased in the order: MCR + CD (27.6%) > PWC (22.8%) > CMS (22.6%) > MS (19.4%). The ready vermicompost obtained from MCR + CD vermibed showed the maximum increase (% of initial level) in content of total N (143.4%), available P (111.1%) and exchangeable K (100.0%). The end product showed reduction in C:N ration between the ranges of 60.7% (CSM) and 70.3% (MCR + CD), at the end. The composting earthworm E. eugeniae exhibited the highest values of biological parameters: maximum mean individual biomass (1261.25 ± 7.0 mg), biomass gain (955.84 ± 11.03 mg), growth rate (10.62 ± 0.10 mg wt. worm⁻¹ day⁻¹), cocoon numbers (87.67 ± 6.51), and reproduction rate (0.66 ± 0.01 cocoons worm⁻¹ day⁻¹) in CSM container, while MS vermibeds showed the lowest values of these parameters. During experimentation, the maximum mortality for E. eugeniae was recorded in MS (16.67 ± 7.63%) followed by CSM > PWC > MCR + CD. Results indicated that the C:N ratio of the substrate drastically influenced the growth parameters of E. eugeniae, and it showed the close relations with maximum individual biomass gain (R² = 0.96), individual growth rate (R² = 0.82), and reproduction rate (cocoon worm⁻¹ day⁻¹) (R² = 0.72), in different treatments. This study clearly indicates that vermicomposting of crop residues and cattle shed wastes can not only produce a value-added product (vermicomposting) but at the same time acts as best culture medium for large-scale production of earthworms.     

Nutrient composition,digestion and rumen fermentation in sheep of wheat straw treated with calcium oxide,sodium hydroxide and alkaline hydrogen peroxide

This study tested the effect of calcium oxide (CaO), sodium hydroxide (NaOH) and NaOH plus hydrogen peroxide (H₂O₂; AHP) on cell wall composition, digestion and fermentation of wheat straw (straw) in sheep. Treated straws were prepared by mixing straw either with water followed by dusting with CaO at 160 g kg⁻¹ DM or with a NaOH solution alone at 3 l kg⁻¹ DM to supply 80 g NaOH kg⁻¹ DM (Na) or pre-soaked with Na exactly as in the previous treatment for 27 h followed by mixing with 130 g H₂O₂ kg⁻¹ DM (AHP) for 6 h. After 14 days of storage, the treated straws and an untreated straw (U) were fed automatically every 2 h to four individually housed sheep together with a supplement in a 4×4 latin square experiment. Each kilogram supplement DM contained 422 g CP and 10.8 MJ ME. NDF (p<0.001) and hemicellulose (p<0.01) contents were significantly reduced whereas cellulose was increased (p<0.001) in treated compared to untreated straw. ADL was reduced in Ca (p<0.05) but increased (p<0.05) in Na and AHP compared with U. The rumen and total tract digestibility were significantly (p<0.001) greater in sheep fed treated compared with untreated straw. Significant differences (p<0.05) between treatments for pH, NH₃ and VFA were also observed. All treatments improved the nutritive value of straws compared with untreated through modification of cell wall with a subsequent increase in digestibility by sheep. Although the digestibility for Ca was lower than that for Na despite reduction in cell wall, its use to treat straws may be more safe and cost effective than Na. AHP was the most effective and could also improve the energy value of other low quality forages for ruminants. However, the need of AHP for a large amount of NaOH to achieve highly alkaline pH limits its farm scale application. Therefore, further studies should either consider reducing the amount of NaOH or finding alternative alkalis that are cost effective and user-friendly.     

Effects of dietary whole cottonseed and crude protein level on rumen protozoal population and fermentation parameters

In this investigation in vitro and in vivo trials were performed to determine the efficacy of a cottonseed to limit protozoal population and fermentation parameters. The composition of diets given to the different treatments were as follow: (1) control (without whole cottonseed), 16% crude protein (CP), 3.2% ether extract (E.E.); (2) 20% whole cottonseed, 16% CP, 6.5% E.E.; (3) 20% whole cottonseed, 13% CP, 6.4% E.E. and (4) 20% crushed whole cottonseed, 13% CP, 6.4% E.E. DM disappearance (DMD) and fermentation characteristics of the treatments were determined by in vitro incubation studies. In the in vivo trial, ruminal fluid was taken by rumenocentesis (3 h after feeding) on days 1, 3, 5, 7, 9, 11, 14, 21 and 28 from four sheep fed about treatment diets. The pH and protozoal counts were determined in each sample, while ammonia nitrogen and volatile fatty acid (VFA) were determined in samples taken on days 7, 14, 21 and 28. The in vitro DMD after 24 h incubation decreased (p < 0.01) with the addition of cottonseed in diets 3 and 4 and DMD after 72 h incubation was highest (p < 0.01) for the control diet. The fractional rate of gas production (c) for the control and diet 2 was higher (p < 0.05) than for the diets 3 and 4. Feeding crushed whole cottonseed decreased molar proportion of propionate (p < 0.05) and increased molar proportion of butyrate (p < 0.01). Low crude protein level increased the molar proportion of propionate (p < 0.05) and decreased molar proportion of butyrate (p < 0.05) and cellolytic protozoa population (p < 0.05). Feeding cottonseed decreased (p < 0.05) the total protozoa population from approximately 500,000 to 250,000 ml⁻¹ and Holotrich and cellulolytic protozoa disappeared from the rumen of sheep and only Entodinium sp., remained. This was associated with lower concentration of ammonia nitrogen in rumen fluid of sheep fed diets 4 (p < 0.05) and 2 (p < 0.01). It was concluded that cottonseed reduced rumen fauna and ammonia nitrogen, but had no effect on ruminal VFA while the crushed whole cottonseed decreased molar proportion of isovalerate only. In vivo molar proportion of propionate and butyrate and valerate were increase and decrease, respectively, by decreasing CP percentage in treatment diets.     

Structure and function of sheep hemoglobin Chios: A novel allele at the HBBB locus with two Lys → Arg substitutions at positions β66(E10) and β144(HC1)

A novel hemoglobin variant was observed in pure sheep (Ovis aries) breeds of the island of Chios (Greece), Egypt and Hungary. This silent variant was identified by gel electrophoresis and RP-HPLC of dissociated globin chains. Two Arg for Lys substitutions were detected, by means of MALDI TOF electrospray mass spectrometric analysis for the intact globins, at positions β66(E10) and β144(HC1) of a globin chain having the sequence of the β^B chain. Sequencing of the β-globin gene confirmed the variant gene as being an allele of the HBBB locus having the AAG → AGG and the AAA → AGA mutations at codons 66 and 144, respectively, both corresponding to the Lys → Arg substitution. The intrinsic oxygen affinity of the variant Hb (logP₅₀ = 0.79 at pH 7.0) was found to be intermediate between that of the sheep Hb B (logP₅₀ = 0.92) and that of Cypriot mouflon (O. a. ophion) Hb (logP₅₀ = 0.53), the latter having only the Lys → Arg change at β144, whereas nearly no differences were observed in the presence of the Cl⁻ physiological effector. Result supports the indication that Arg at β144 enhances the role of the ligand in decreasing oxygen affinity, this effect being partially counteracted when Arg is at β66. Data also shows that the Lys → Arg change at β66 is responsible for 1.49 fold reduction in the intrinsic oxygen affinity. This hitherto undescribed variant increases to seven the number of alleles at the sheep HBBB locus. Following the nomenclature used for human Hb variants, the new allele was termed as the Hb Chios or [β^B66(E10) Lys → Arg, 144(HC1)Lys → Arg], whereas the proposed genetic nomenclature of the locus is HBBK.