Organic zinc and copper supplementation on antioxidant protective mechanism and their correlation with sperm functional characteristics in goats

Trace minerals feeding had significant effects on sperm production and fertility with better absorption and proper utilization within the body for optimum reproductive function. Several studies have shown that more influenced trace elements in the diets of animals are copper (Cu) and zinc (Zn). Bucks showing deficiency of this mineral might affect the quality of semen production which in turn would affect the fertility. This experiment was thus designed to test the effects of organic Cu and Zn supplementation on antioxidants enzyme activities and sperm functional attributes in fresh semen of bucks. Forty bucks (n = 40, Aged 5 months) were assigned to ten groups of four animals in each group, supplemented (for a period of 8 months) with different levels of organic Zn: 20 mg (T2), 40 mg (T3) and 60 mg (T4), organic Cu: 12.5 mg (T5), 25 mg (T6), 37.5 mg (T7) and combined organic Zn and Cu: 20 + 12.5 mg (T8), 40 + 25 mg (T9), 60 + 37.5 mg (T10), respectively, per kg dry matter and no additional mineral diet (control; T1). One hundred and sixty semen samples were collected through electro‐ejaculator and analysed for sperm quantity, quality, acrosome intactness and plasma membrane integrity and correlated with the catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzyme activities in seminal plasma. The results indicated organic Cu and zinc supplemented bucks produced more sperm cells, had higher sperm concentrations, maintained higher (p < .01) sperm livability, plasma membrane and acrosome integrities, more motility and velocity. The increased antioxidant enzyme activities, reduced oxidative stress and lowered lipid peroxidation were positively correlated (p < .05) with the sperm functional attributes. In conclusion, organic Cu and Zn supplement to male goats showed protective roles against oxidative damage and maintained better fresh semen characteristics.     

The effects of health classification and housing and management of feeder pigs on performance and meat inspection findings of all-in-all-out swine-finishing herds

The effects of health classification and of housing and management on performance and meat inspection findings were studied in 166 all-in-all-out finishing herds in Finland in 1995 and 1998. Producers could buy either health-classified (certified free of certain diseases) or standard-class pigs for their farms. Herds that had attained a certain level of housing and management were eligible to join the LSO 2000 management system for finishing herds. In 1995, none of the study herds were LSO 2000 units. In 1998, 76 of them had become LSO 2000 units, while 90 had remained as non-LSO 2000 units. Continuous outcome variables (daily gain and time in the finishing unit) were analysed with a mixed-model procedure with repeated measurements from the same farms. Discrete variables were analysed either with Poisson regression (mortality, whole- and partial-carcass condemnations, organ condemnations, arthritis, abscesses) or logistic regression (liver condemnations, pneumonia, pleuritis). The models were adjusted for the year, size of the herd and the average slaughter weight of the batch.The benefits obtained by the health classification of the feeder pigs included a substantial increase in daily gain (37g, about 5%) and decrease in time in the finishing unit (4 days, about 4%), decrease in mortality (odds ratio, OR 0.68), a substantial decrease in pneumonia (OR 0.37) and pleuritis (OR 0.60), and a substantial decrease in liver (OR 0.45) and organ (OR 0.72) condemnations. Classification of the pigs did not have an effect on the prevalence of whole-carcass condemnations, arthritis or abscesses. The health-classified pigs had, unexpectedly, more partial-carcass condemnations (OR 1.15) than the standard-class pigs. The obtained benefits of the LSO 2000 units were an increase in daily gain (14g, about 2%), decrease in time in the finishing unit (2 days, about 2%), a decrease in mortality (OR 0.69) and a decrease in whole-carcass (OR 0.69) and partial-carcass (OR 0.81) condemnations. The LSO 2000 units did not have any advantage over the non-LSO 2000 units in their prevalence of liver or organ condemnations, arthritis, abscesses, pneumonia or pleuritis.     

Evaluation of fluorochromes for imaging bacteria in soil

The distribution of inoculated Escherichia coli cells and indigenous bacteria in a silt loam soil and a sandy soil was studied using fluorescence microscopy techniques. Ethidium bromide stained inoculated cells against the soil and resin background. Satisfactory results were also obtained with epifluorescence microscopy of samples stained with calcofluor white M2R (CFW) and 4′,6-diamidino-2-phenylindole (DAPI). Indigenous soil bacteria were visualized in soil thin sections, after staining with ethidium bromide, using fluorescence microscopy. The density of these bacteria was estimated to be 10⁷-10⁸ cells/cm³. Inoculated E. coli cells, stained with one of the green fluorochromes (fluorescein isothiocyanate (FITC), 5-(4,6-dichlorotriazinyl)aminofluorescein, or eosin Y), could be clearly distinguished in sandy soil thin sections. Confocal laser scanning microscopy with 3D image reconstruction was also successfully applied to characterize distribution of E. coli introduced to soil. The fate of introduced bacteria and the location of indigenous bacteria in soil can be confirmed using the microscopic techniques described in this paper.     

Modelling and Mapping Ozone Deposition in Europe

A new dry deposition module has been developed for European-scale mapping and modelling of ozone deposition fluxes (Emberson et al., 2000a,b). The module is being implemented in the photochemical long-range transport model of EMEP that is currently used to estimate exceedance of the existing critical levels for ozone within the UN ECE LRTAP programme. The deposition model evaluates the atmospheric, boundary layer and surface resistances to ozone transfer with the calculation of the dry deposition velocity performed according to a standard resistance formulation. The approach differs from other existing methods through the use of a detailed stomatal uptake model that describes stomatal conductance as a function of plant species, phenology and four environmental variables (air temperature, solar radiation, water vapour pressure deficit and soil moisture deficit). Comparison of preliminary model outputs for selected land-cover types indicate that the model is capable of predicting the seasonal and diurnal range in deposition velocities that have been reported previously in the literature. The application of this deposition scheme enables calculations of ambient ozone concentrations to be made using a biologically based method that can distinguish stomatal and non-stomatal components of total ozone deposition. The ability to estimate stomatal ozone fluxes (according to vegetation type, phenology and spatial location) that are consistent with evaluations of atmospheric ozone concentrations will be helpful in future assessments of ozone impacts to vegetation.     

Biomineralization of atrazine and analysis of 16S rRNA and catabolic genes of atrazine degraders in a former pesticide mixing site and a machinery washing area

Purpose

The purpose of this study was to determine the first-order rate constants and half-lives of aerobic and anaerobic biomineralization of atrazine in soil samples from an agricultural farm site that had been previously used for mixing pesticide formulations and washing application equipment. Atrazine catabolic genes and atrazine-degrading bacteria in the soil samples were analyzed by molecular methods.

Materials and methods

Biomineralization of atrazine was measured in soil samples with a [U-ring-¹⁴C]-atrazine biometer technique in soil samples. Enrichment cultures growing with atrazine were derived from soil samples and they were analyzed for bacterial diversity by constructing 16S rDNA clone libraries and sequencing. Bacterial isolates were also obtained and they were screened for atrazine catabolic genes.

Results and discussion

The soils contained active atrazine-metabolizing microbial communities and both aerobic and anaerobic biomineralization of [U-ring-¹⁴C]-atrazine to ¹⁴CO₂ was demonstrated. In contrast to aerobic incubations, anaerobic biometers displayed considerable differences in the kinetics of atrazine mineralization between duplicates. Sequence analysis of 16S rDNA clone libraries constructed from the enrichment cultures revealed a preponderance of Variovorax spp. (51 %) and Schlesneria (16 %). Analysis of 16S rRNA gene sequences from pure cultures (n?=?12) isolated from enrichment cultures yielded almost exclusively Arthrobacter spp. (83 %; 10/12 isolates). PCR screening of pure culture isolates for atrazine catabolic genes detected atzB, atzC, trzD, trzN, and possibly atzA. The presence of a complete metabolic pathway was not demonstrated by the amplification of catabolic genes among these isolates.

Conclusions

The soils contained active atrazine-metabolizing microbial communities. The anaerobic biometer data showed variable response of atrazine biomineralization to external electron acceptor conditions. Partial pathways are inevitable in soil microbial communities, with metabolites linking into other catabolic and assimilative pathways of carbon and nitrogen. There was no evidence for the complete set of functional genes of the known pathways of atrazine biomineralization among the isolates.

     

Significance of earthworms in stimulating soil microbial activity

 The stimulatory effect of earthworms (Lumbricus terrestris L.) on soil microbial activity was studied under microcosm-controlled conditions. The hypothesis was tested that microbial stimulation observed in the presence of a soil invertebrate would be due to the utilization of additional nutritive substances (secretion and excretion products) that it provides. Changes in microbial activity were monitored by measuring simultaneously CO₂ release and protozoan population density. The increase in CO₂ released in the presence of earthworms was found to result from both earthworm respiration and enhanced microbial respiration. The stimulation of microbial activity was confirmed by a significant increase in protozoan population density, which was 3–19 times greater in the presence of earthworms. The respiratory rate of L. terrestris was estimated to be 53 μl O₂ g^–1 h^–1. Earthworm respiration significantly correlated with individual earthworm weight, but there was no correlation between the increase in microbial respiration and earthworm weight. This finding does not support the hypothesis given above that enhanced microbial respiration is due to utilization of earthworm excreta. A new hypothesis that relationships between microbial activity and earthworms are not based on trophic links alone but also on catalytic mechanisms is proposed and discussed. Received: 26 August 1997     

Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.     

Bacterial phylogenetic diversity in a constructed wetland system treating acid coal mine drainage

Microorganisms in acid mine drainage are typically acidophiles that mediate the oxidation of reduced compounds of iron and sulfur. However, microbial populations in wetland systems constructed to treat acid mine drainage are not well characterized. The purpose of this study was to analyze bacterial diversity, using cultivation-independent molecular ecological techniques, in a constructed wetland that received acid drainage from an abandoned underground coal mine. DNA was purified from Fe(III)-precipitates from the oxidized surface zone of wetland sediments and 16S rRNA gene sequences were amplified and cloned. A total of 200 clones were analyzed by restriction fragment length polymorphism (RFLP) and 77 unique RFLP patterns were obtained with four restriction enzymes. Of these patterns, 30 most dominant unique clones were selected for sequencing of their 16S rRNA genes. Half of these 30 clones could be matched with autotrophic iron- and sulfur-oxidizing bacteria (Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans). Several clones also formed a clade with heterotrophic iron-oxidizing bacteria (TRA2-10, TRA3-20, and TRA5-3) and heterotrophic bacteria (Stenotrophomas maltophilia, Bordetella spp., Alcaligenes sp., Alcaligenes faecalis, and Alcaligenes xylosoxidans). Approximately 40% and 35% of the analyzed RFLP restriction patterns were consistent with A. ferrooxidans and A. thiooxidans, respectively. The relatively high frequency of acidithiobacilli is consistent with the chemical and physical characteristics of this site—i.e., continuous, abundant supply of reduced iron and sulfur compounds, pH 3–4, ambient temperature, and limited organics originating from the coal seam and from vegetation or soil surrounding the inlet channel to the wetland. The RFLP results were consistent with our previous culture-independent PCR-DGGE and FISH study, showing relatively low bacterial diversity and predominance of mesophilic acidithiobacilli in oxic wetland sediments.     

Diagnoses and treatments in health-classified fattening herds rearing pigs all in-all out

This study describes diseases encountered, medications used and veterinary involvement in all in--all out finishing herds belonging to one pork production system. The finishing herds had a particular management and housing regime. The pigs originated from health classified farrowing units. Information on 207,442 pigs was collected from 595 log books. Altogether 91% of the pigs received no treatments. Four percent of the batches of pigs were given antimicrobial mass medications. The local veterinarian visited the herds on average 2.6 times during the finishing period and made the diagnoses in more than half of the cases. At least one pig was affected with arthritis or tail biting in more than half of the batches, whereas locomotory diseases were recorded in one third of the batches. All other diagnoses were encountered in 1%-13% of the batches. Only a few pigs were treated individually in the affected groups. Antimicrobial drugs were given to 8% and other medicines to 0.7% of the pigs. The diagnosis was missing at least for one pig in 29% of the batches and the information about the medicine use in 8% of the treatments was missing. The study shows that it is possible to rear finishing pigs with only a small proportion of the animals needing treatments. The need of mass medications was low, because infectious diseases affecting the whole herd were uncommon. The recommendations for antimicrobial use given by the authorities had been followed quite well. The farmers and the veterinarians should be educated in order to realise the importance of proper record keeping.     

Detectability and prevalence of Brachyspira species in herds rearing health class feeder pigs in Finland

Faeces samples were taken three times at two-week intervals, from the farrowing units of four herds of known Brachyspira (formerly Serpulina) status and one of unknown Brachyspira status. Brachyspira hyodysenteriae, Brachyspira pilosicoli, Brachyspira intermedia and Brachyspira group III were isolated from the faecal samples from the weaners in the herds using either a maximum of 50 ppm of olaquindox or no feed additives. The detection rates were relatively consistent. However, B hyodysenteriae was not detected at one sampling in a known positive herd. The prevalence of Brachyspira species was also studied in feeder pigs originating from LSO 2000 health class farrowing units, comparable with specific pathogen-free herds. These farms were free from swine dysentery, sarcoptic mange, swine enzootic pneumonia and progressive atrophic rhinitis. Fifty of 428 herds were sampled once. B hyodysenteriae was not isolated from any of them, but B intermedia, B pilosicoli and Brachyspira group III were isolated from five, 14 and 37 of the herds, respectively. The detection of Brachyspira species did not relate to the prevalence of diarrhoea in the herds, as judged by the farmers. The herds using carbadox (40 to 50 ppm) had a lower prevalence of Brachyspira species than those using olaquindox (40 to 50 ppm).