Association of Mycobacterium avium subspecies paratuberculosis infection with milk production and calving interval in Iranian Holsteins

There are inconsistent results for the association of Mycobacterium avium subspecies paratuberculosis (MAP) infection with production and reproduction in dairy cows. Determination of these associations in each region is essential to encourage participation of dairy cattle producers in disease control programs. This study was conducted in Shiraz, southern Iran, to quantify the association of subclinical MAP infection with 305-day milk production and calving interval in Iranian Holsteins. A total of 21 dairy herds were selected for the study and in each herd, quarter milk samples were collected from ten to 12 dairy cows for PCR analysis. Data about parity, calving interval, length of lactation period, total milk production and 305-day milk production were also provided for each animal. Overall, 252 individual milk samples were collected. Herd- and individual-level prevalence of MAP infection were 23.8% (95% CI, 6.2–41.4%) and 3.2% (95% CI, 1.3–5.1%), respectively based on IS900 nested PCR. The results for 305-day milk production revealed a 248 kg reduction in positive cows compared with negative ones (P = 0.009). When cows from positive herds were compared with cows from negative herds, a 335-kg reduction in 305-day milk production (P = 0.005) and a 30-day increase in calving interval (P = 0.057) were observed in the former group. These findings support the previous results that paratuberculosis infection is negatively associated with the performance of the animals.     

Immunohistochemical and polymerase chain reaction studies in Neospora caninum experimentally infected broiler chicken embryonated eggs

The diagnostic characteristics of immunohistochemistry (IHC) and polymerase chain reaction (PCR) methods were studied in the tissues of broiler chicken embryos experimentally infected by Neospora caninum. An infection with N. caninum NC-1 isolate was conducted in 70 broiler chicken embryonated eggs randomly divided into seven equal groups. After 8 days of incubation, six groups were inoculated with 10, 10(2), 10(3), 10(4), 10(5), and 10(6) doses of tachyzoites/embryonated egg. The 7th group was considered as control. The mortality rate and pathological changes of the dead embryos and hatched chickens up to 60 days old were noticed. Consecutive sections to those used for histopathological examination including the liver, heart, brain, and chorioalantoic (CA) membrane were subjected to IHC. The intensity and distribution of the immunostaining was graded as highly to mildly positive. For PCR procedure, DNA was extracted from 50mg of the tissues and primer pair Np21/Np6 was used for amplification of the Nc-5 gene. The results of the immunosignaling ranged from variable degrees of mild to moderate staining as dark-brown to brown and coarsely to finely granular, mostly within the cytoplasm of infected cells such as the endothelial cells of blood vessels. The parasite aggregation was more predominant in the heart than other tissues. Immunoreactivity for N. caninum antigen was multifocally moderate positive in the heart, liver and CA of the 10(3) dose, and also heart, liver, brain and CA of the 10(4) dose. IHC showed mildly positive in the liver and heart of the chicken embryos infected with 10 and 10(2) tachyzoites, as well. The results of the PCR confirmed the existence of the parasite in all of the examined tissues from the 10(3) and 10(4) doses. In conclusion, the results indicate a good agreement between IHC and PCR in diagnosis of neospora antigen in the infected tissues.     

Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It is native to Iran and spread from Iran to other areas. In this study amplified fragment length polymorphism (AFLP) was used to detect intra- and inter-population genetic diversity of pomegranate. A group of 67 accessions belonged to 4 populations from Iran was studied using eight primer combinations. A total of 221 scorable bands were amplified, of which, 118 (54.13%) were polymorphic. Resolving power (Rp) ranged from 5.70 to 9.21, and the average of polymorphism information content (PIC) per primer pair was 0.40. According to Nei's gene diversity and allelic statistics, Isfahan population had a highest genetic diversity (H = 0.3646, I = 0.5327, N_e = 1.6467). Coefficient of gene differentiation between populations (GST) was 0.124, indicated that mainly proportion of genetic variation (87.6%), was within populations and the remaining (12.4%) of the variation was among populations that, also supported by analysis of molecular variance (AMOVA). The gene flow (Nm) varied from 0.969 to 10.404 between pair-wise populations and was 3.504 among all of the populations. The Jaccard similarity coefficient between individuals ranged from 0.26 to 0.88. The UPGMA dendrogram clustered all 67 accessions into 6 groups. In some cases accessions from same region were grouped together but in most cases, there was gene exchange. To study the genetic relationships among populations, a principal coordinate analysis (PCoA) based on Nei's genetic distances was performed. Results of this study showed that AFLP marker can be a useful tool for investigating the genetic diversity of pomegranate genotypes.     

Innate immune defense system of the skin

Background – Antimicrobial peptides (AMPs) have a pivotal role in cutaneous innate immunity. They are present in the skin of many animals, including mammals, and are both constitutively present and inducible by infection and injury. Functions – Antimicrobial peptides exhibit antimicrobial activity against bacteria, viruses, fungi and parasites, with different potencies depending on their peptide structure. They also act as multifunctional effector molecules that influence diverse cellular processes, including cell migration, proliferation and differentiation, cytokine production, angiogenesis and wound healing. Suppressed AMP production has been associated with increased susceptibility to microbial insults and the pathogenesis of atopic dermatitis. This review highlights recent observations on the expression and role of AMPs, particularly the AMPs cathelicidin and β‐defensin, in healthy and diseased skin.     

Role of selenium in mitigation of cadmium toxicity in pepper grown in hydroponic condition

Selenium (Se) is an essential element for humans but is not considered as essential for plants. However, its beneficial role in improving plant growth and stress tolerances is well established. In order to study the role of Se in cadmium (Cd) toxicity in pepper (Capsicum frutescens cv. Suryankhi Cluster), this experiment was carried out in greenhouse conditions. Treatments comprised Cd [0, 0.25, and 0.5 mM cadmium chloride (CdCl₂)] and Se [0, 3, and 7 µM sodium selenite (Na₂SeO₃)] with three replications. The result showed that Cd decreased chlorophyll a, chlorophyll b, and carotenoids, whereas Se supplementation diminished Cd toxicity on photosynthetic pigment. Selenium at 7 µM significantly increased the leaf area in the plants grown at 0.25 mM Cd. The application of Se at 3 µM with 0.25 mM Cd and Se at 3 µM and Se at 7 µM with 0.5 mM Cd increased the activity of catalase (CAT). Selenium at 7 µM decreased the proline content of pepper leaves exposed to Cd at 0.5 mM (30%). Selenium significantly enhanced the antioxidant activity of leaves, which was diminished by Cd toxicity. In general, Se has a beneficial effect on plant growth and is an antioxidant enzyme of pepper cv. Suryankhi Cluster under Cd stress and non-stress conditions.     

Effects of drought stress on the seedling growth,development, and metabolic activity in different cultivars of canola

ABSTRACT

The comparative effects of drought stress on seed performance, growth parameters, free proline content, lipid peroxidation, and several antioxidative enzymes activities were studied in both cultivars of Brassica napus L. Drought stress increased mean germination rate and mean of day germination in both cultivars, and its effect was more pronounced in RGS003. The length and dry weight of root increased significantly in both cultivars under drought. RGS003 was more tolerant and obtained more biomass under drought than that of Sarigol. Proline content and pyrroline-5-carboxylate synthetase expression increased in Sarigol under drought. Measurement of malondialdehyde content in seedlings showed that lipid peroxidation was lower in RGS003 than in Sarigol. Antioxidant enzyme activities showed different trends in the two cultivars under drought stress but were higher in RGS003 than in Sarigol. These results suggest that RGS003 is better protected against drought-induced oxidative damage. Lipoxygenase activity only induced under water deficit condition in RGS003. Changes of respiratory enzymes activities of RGS003 subjected to drought stress showed a pattern different with Sarigol. Drought stress induced aconitase activity in RGS003, but it reduced fumarase and succinate dehydrogenase activity in Sarigol. This study showed that RGS003 exhibits a better protection mechanism against oxidative damage and RGS003 is more drought-tolerant than Sarigol possibly by maintaining and/or increasing growth parameters, antioxidant enzyme activities, and respiratory enzymes activities.     

The frequency of eight common point mutations in CYP21 gene in Iranian patients with congenital adrenal hyperplasia

BACKGROUND: Congenital Adrenal Hyperplasia (CAH, the inherited inability to synthesize cortisol) is one of the most common (1 in 10000 to 1 in 15000) autosomal recessive disorders. More than 95% of cases of CAH are caused by 21-hydroxylase deficiency (21-OHD). Females with severe, classic 21-OHD are exposed to excess androgens prenatally and are born with virilized external genitalia. Most patients cannot synthesize sufficient aldosterone to maintain sodium balance and may develop potentially fatal salt wasting crisis if not treated. METHODS: We applied allele specific PCR to detect the eight common mutations in the CYP21 gene in patients. Fifty unrelated patients with symptoms of classical CAH were studied. RESULTS AND CONCLUSION: Seventy percent of our subjects had these mutations. The most frequent mutations were found to be I2G and del-8 bp (28% and 13%, respectively). The frequencies of other alleles were as following: I172N, 9%; V281L, 3%; exon 6 cluster (I236N, V237E and M239K), 4%; Q318X, 9%; R356W, 5%; and P30L, 0%. The frequency of mutations did not differ substantially from other ethnics, however, a higher rate of del-8 bp (13%) was found in our population. The aim of this study was to detect common mutations for setting up a molecular method for prenatal diagnosis.     

Phylogenetic relationship and genetic differentiation of Populus caspica and Populus alba using cpDNA and ITS noncoding sequences

Populus caspica Bornm.(section Leuce and subsection Albida), one of the most endangered endemic tree species in the Hyrcanian Forest in Iran, has numerous morphological characteristics that are closely similar to Populus alba; to clarify their taxonomic relatedness and genetic differentiation and thus inform conservation strategies, we used the noncoding regions of chloroplast DNA(cpDNA; trnL-F and trnH-psbA) and the internal transcribed spacer(ITS). Leaf samples were collected from six populations across northern Iran. cpDNA and ITS fragments were amplified by universal primers using the PCR technique and directed sequencing. The results showed that P. caspica is genetically differentiated from P.alba, and two ITS variants were detected within some P.caspica individuals. Conflicts between topologies from ITS and plastid genomes were observed. High differentiation of P. caspica from the other Populus species shown in this study confirmed the diverging taxonomic status of this endangered species. We recommend in situ conservation measures(e.g., protected areas) for at least several populations of this species, especially in the plain regions of the Hyrcanian forest.