Morphometric study of the testis in sheep embryos using unbiased design-based stereology

Sheep have been used as translational models of human postnatal testicular development. However, the morphometric features of the normal developing testis in sheep embryos have not been previously investigated using stereology. The objective of the present work was to establish normal quantitative parameters for fetal testicular tissue components in sheep, using unbiased design-based stereological methods. Twenty-four sheep embryos were divided into four gestational age groups (9–11, 12–14, 15–17 and 18–20 weeks of gestation) on the basis of the embryos’ crown-rump length. Isotropic, systematic uniform random sections of the left testes were obtained by employing the orientator method. Testicular total volume, the absolute and proportional volumes occupied by the seminiferous tubules and interstitial tissue, as well as the seminiferous tubule length, were estimated using the point-counting system and the unbiased counting frame principle. All the parameters, with the exception of the interstitial tissue's fractional volume, gradually increased along with gestational age, with the maximum increase especially seen in the late fetal stages. The proportional volume of the interstitial tissue, on the other hand, showed a decreasing trend along with increasing gestational age. The absolute volume of the testes, of the seminiferous tubules and of the interstitial tissue, and the length of the seminiferous tubules showed a significant (p< 0.05) positive linear correlation with gestational age. Several similarities were observed with human testicular embryogenesis. The stereological data emerging from the present study might prove useful as basic contribution to the fields of andrology and embryology and stimulate further research in these areas.     

Determination of antibiotic resistance pattern and virulence genes in Escherichia coli isolated from bovine with subclinical mastitis in southwest of Iran

The aims of the present study were to investigate the prevalence of some virulence genes and also determine the antimicrobial resistance pattern of E. coli isolated from bovine with subclinical mastitis. The milk of 502 cows was collected from 8 dairy herds in the southwest of Iran. Conventional biochemical tests were used for identification of E. coli at the species level. Antimicrobial susceptibility patterns of E. coli isolates were determined by disc agar diffusion method and polymerase chain reaction (PCR) was used for detection of seven virulence genes including f17A, afaE-8, afaD-8, eaeA, cnf1, cnf2, and iucD. Seventy (13.94%) isolates of E. coli were identified in 502 milk samples. The highest rate of resistance was observed against tetracycline (18.6%), while none of the isolates were resistant to streptomycin. Eight (11.5%) out of 70 E. coli isolates carried at least one of the virulence genes. The afaD-8 was the most prevalent gene detected in 5 (7.1%) isolates. The afaE-8, iucD, and eaeA were detected in 3, 3, and 2 isolates respectively. Low prevalence of virulence factors may be indicating that most of the E. coli isolates originated from the commensal flora of cows and enter to the udders via environment contamination with feces.

     

Bacterial DNA Detection in the Blood of Healthy Subjects

Background:The presence of microbiome in the blood samples of healthy individuals has been addressed. However, no information can be found on the healthy human blood microbiome of Iranian subjects. The current study is thus aimed to investigate the existence of bacteria or bacterial DNA in healthy individuals. Methods: Blood samples of healthy subjects were incubated in BHI broth at 37 °C for 72 h. The 16S rRNA PCR and sequencing were performed to analyze bacterial isolates. The 16S rRNA PCR was directly carried out on DNA samples extracted from the blood of healthy individuals. NGS was conducted on blood samples with culture-positive results.Results:Fifty blood samples were tested, and six samples were positive by culture as confirmed by Gram staining and microscopy. The obtained 16S rRNA sequences of cultured bacterial isolates revealed the presence of Bacilli and Staphylococcus species by clustering in the GeneBank database (≥97% identity). The 16S rRNA gene sequencing results of one non-cultured blood specimen showed the presence of Burkholderia. NGS results illustrated the presence of Romboutsia, Lactobacillus, Streptococcus, Bacteroides, and Staphylococcus in the blood samples of positive cultures.Conclusion: The dormant blood microbiome of healthy individuals may give the idea that the steady transfer of bacteria into the blood does not necessarily lead to sepsis. However, the origins and identities of blood-associated bacterial rDNA sequences need more evaluation in the healthy population.Key Words: Bacteria, Blood, Microbiome, Sequencing, 16S rRNA     

Survey of the microbiological count of the milk collected from livestock ancient city of Garmsar,Semnan Province,Iran

Milk has a high nutritional value and can affect human health. Therefore, it is important to know the factors affecting its quality. The purpose of this study was to evaluate the raw milk microbial quality and factors affecting it. A total of 4260 raw cow milk samples, which were collected from villages around Garmsar city, Iran, and their microbial load were registered at Garmsar Dairy Cow Association between 2013 and 2014, was used in the present study. The total average ± SD of microbial load concentration was 3.02?×?10⁵?±?2.75?×?10⁵ (CFU/ml). The average concentration of total bacteria in 2013 was 2.73?×?10⁵?±?1.58?×?10⁵ (CFU/ml) and in 2014 it was 3.32?×?10⁵?±?3.93?×?10⁵(CFU/ml). Seasonally, it was 2.94?×?10⁵?±?1.72?×?10⁵ (CFU/ml), 3.69?×?10⁵?±?4.55?×?10⁵ (CFU/ml), 2.91?×?10⁵?±?1.62?×?10⁵ (CFU/ml), and 2.63?×?10⁵?±?1.67?×?10⁵ (CFU/ml) for spring, summer, autumn, and winter, respectively. There was a significant difference in seasonal average microbial concentration of milk samples (p?<?0.05). The present study concludes that microbial load of milk can be affected by environmental and hygienic conditions. Ranchers need to pay more attention to hygiene especially in warm seasons. Because in case of high microbial load in raw milk, even after milk pasteurization, it is not possible to have a high-quality and desirable product. Milk producers and distributors need also awareness in order to reduce the incidence of secondary contamination. But, the main correction is to establish direct supervision of the Ministry of Health onto the health of dairy cow farmers by structural integrity of Veterinary Medical Organization as the global one health.

     

Genetic diversity and population structure of four Iranian alfalfa populations revealed by sequence-related amplified polymorphism (SRAP) markers

In the present study, genetic diversity of 48 individual plants from four Iranian cultivated populations of alfalfa (Medicago sativa L.) was evaluated using sequence-related amplified polymorphism (SRAP) markers. Fourteen SRAP primer combinations produced 193 fragments of which 95 were polymorphic. The number of polymorphic fragments detected per primer combination ranged from 3 to 10 bands with an average of 6.78 bands. Average polymorphic information content (PIC) was 0.343 for all primer combinations. Although the AMOVA (analysis of molecular variance) results showed a significant difference in the genetic diversity among the populations (P < 0.0001), the genetic variation mainly caused by the variation of intra population accounted for 93.17% of the total genetic variation. Unweighted pair-group method arithmetic average (UPGMA) analysis of the marker data clearly separated the populations of subtropical (Yazdi) and semi-cold (Hamadani and Nikshahri) as well as Kodi, an improved population. It can be concluded that SRAP markers are useful for studying diversity and relationships among and within alfalfa populations.     

Changes in essential oil accumulation of Moldavian balm (Dracocephalum moldavica L.) in response to phosphate biological and chemical fertilizer

A field experiment was carried out with a split plot arrangement based on a randomized complete block design with three replications in 2014. Triple super phosphate (0, 3.75, 7.5, 11.25 and 15 g m^?2) was applied in main plots and phosphate bio-fertilizer (seed inoculation and topdressing) was allocated to sub plots. Results indicated that application of plants with phosphorus (P) bio-fertilizer as topdressing increased plant height and the number of the lateral branches under low levels of chemical P fertilizer, while plants from seed inoculated with P bio-fertilizer had more of those traits under high levels of chemical P fertilizer. Maximum leaf and flower dry weights were found under the lowest level of chemical P fertilizer. Moldavian balm plant demonstrated highly responsiveness to biological fertilizers in terms of increase in essential oil (EO) percentage and yield. It can be concluded that the effects of bio-fertilizer as seed inoculation is more during the early vegetative growth of plants. Thus, plants from seeds inoculated with P bio-fertilizer treatment had more biomass and EO percentage under high levels of chemical P fertilizer.     

A methodology for up-scaling irrigation losses

This paper presents a methodology for up-scaling field irrigation losses and quantifying relative losses at the irrigation area level for potential water savings. Two levels of analysis were considered: First, the field level where irrigation is applied. Second, the irrigation area level, where the field level losses are aggregated, or up-scaled, using average loss functions. In this up-scaling approach, detailed crop-soil-water modelling can capture the variability of physical parameters (such as soils, crops, water table depth, and management practices) at the field level which are then used to derive loss functions for aggregating losses at higher scales (irrigation area level). This allows potential field-level adaptations and water management changes made by individual farmers to be assessed for impact at the larger irrigation area level. The APSIM farming systems model was used for simulation of crops (wheat, rice, and soybean) and their interaction with the wider system processes at the field level. Given the climate, soil, and management information (sowing, fertilisation, irrigation, and residue management), the model simulates infiltration, the soil moisture profile, plant water uptake, soil evaporation, and deep drainage on a daily basis. Then, by placing the field level analysis in the context of the wider irrigation system or catchment, it is possible to correlate field level interventions (e.g. water savings measures) with water requirements at these higher levels. Application of this method in the Coleambally Irrigation Area in NSW, Australia, demonstrated that an exponential function can describe the relationship between deep drainage losses and the water table depth for different soil, crop, and water table depth combinations. The rate of loss increase (slope of the curve) with the water table depth is higher on lighter (higher intake rates) soils than on heavy soils and is more pronounced in areas under rice cultivation. We also demonstrate that this analysis technique can assist in identifying spatial distribution of losses in irrigation areas, considering water table depth as an additional factor, leading to targeted areas for water-saving measures.     

NEW MOLYBDENUM-HYPERACCUMULATOR AMONG PLANT SPECIES GROWING ON MOLYBDENUM MINE- A BIOCHEMICAL STUDY ON TOLERANCE MECHANISM AGAINST METAL TOXICITY

The aim of this work was to determine metal accumulation by plants growing on three molybdenum-mine zones and their tolerance strategies. The plants from tailing, extracting and non-contaminated zones were sampled with their corresponding soils. The results show that molybdenum (Mo) and copper (Cu) were at toxic levels in soils and their levels varied in 44 collected species from 21 families. Ajuga chamaecistus and Cramb orientalis L. excluded Mo and Cu, respectively. Achilla tenuifollia as Mo-hyperaccumulator with total Mo (1979 mg kg^?1) and then Erodium ciconium with 1308 mg kg^?1 Mo and Conyza Canadensis with 618 mg kg^?1 Cu were moderate metal accumulators. They stored considerable levels of metals in their leaves vacuoles and elevated the levels of phytochelatins, cysteine and glutathione and induced antioxidant enzymes. In conclusion, this study indicated that some collected plants excluded metals. In metal-accumulators, antioxidant enzymes, phytochelatins and sequestration of excess metals were involved in their tolerance mechanism.     

Development of a monoclonal blocking ELISA for the detection of antibody to Mycoplasma bovis in dairy cattle and comparison to detection by PCR

A monoclonal antibody blocking enzyme-linked immunosorbent assay (B-ELISA) was developed to detect antibodies to Mycoplasma bovis in cattle sera. The assay was highly specific and sensitive and there was no cross-reaction detected. This method revealed a high prevalence of antibodies (60%) to M. bovis in dairy cattle in North Queensland. The diagnostic potential of this B-ELISA for the detection of antibody to M. bovis was compared with its detection by PCR. There was a strong positive correlation between PCR and B-ELISA titers. Thus, the B-ELISA appears to be a valuable and reproducible tool in the serodiagnosis of M. bovis infection in cattle.     

Suppression of telomerase activity by pyrimethamine: implication to cancer

BACKGROUND: Although pyrimethamine (Tindurin) appears to be effective in the prevention and treatment of some infectious diseases, very little information exists on its unpredictable properties. We design this study to evaluate its anti-tumoral effect on a model of cell line. METHODS: The cytotoxic influence of Pyrimethamine on prostate cell line was investigated using an in vitro colometric assay. The potential modulatory effects on metastasis, apoptosis, and immortality characteristics of cells were assessed with gelatin zymography, terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay and telomeric repeat amplification protocol, respectively. RESULTS: Cytotoxicity analysis of pyrimethamine revealed a dose-dependent fashion. An apoptotic influence of pyrimethamine was also confirmed by data obtained from TUNEL assay. Dose-dependent inhibitory effect on matrix metalloproteinases (MMP) was seen in pyrimethamine. A potent inhibitory effect of pyrimethamine was also established by data achieved from TRAPeze telomerase detection kit. CONCLUSIONS: Collectively, as induction of apoptosis together with MMP and telomerase inhibition could be indicative of cancer treatment, pyrimethamine might be considered as a chemopreventative agent in cancer.