Background: Theileria Spp. is the causative agent of the economically significant hemoprotozoan disease known as theileriosis in cattle. In dairy animals, it results in death and decreased productivity. This study aimed to clarify the effects of Theileria annulata (T. annulata) infection on the histological and haemato-biochemical parameters in infected cattle. 
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Methods: Between December 2022 and May 2023, 200 cattle were randomly selected from the slaughterhouse and blood samples were collected from them to assess the hemato-biochemical parameters and screen for T. annulata infection using a Giemsa-stained blood smear, furthermore, the histological profile of the spleen, lymph nodes and liver. 
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Result: T. annulata was detected in 26.5% of the cattle based on blood smear microscopy. The haematological analysis showed a notable increase in lymphocytes and a significant drop in platelets, haemoglobin, total leukocyte count and total erythrocyte count. In cattle infected with T. annulata, biochemical examination showed a significant increase in alanine aminotransferase (ALT), aspartate aminotransferase (AST), high density lipoprotein (HDL) and triglyceride, also increase in bilirubin. Histological examination revealed necrotic degenerations of splenic nodules, oedema, parenchymal necrotic degeneration, increased intercellular spaces, depletion of lymphocytes, increased sinuses in lymph nodes and focal necrosis of hepatocytes with infiltration of inflammatory cells and vacuolar degeneration of hepatocytes.

Impact of Theileria annulata on Hemato-biochemical and Histological Parameters in Cattle

<div>Background: Theileria annulata is predominant and of utmost economical importance tick borne pathogen of bovines in the region which is routinely diagnosed based on the microscopic examination of Romanowsky stained thin blood smears. Present study was intended to evaluate and analyze the detection efficacy of a commercial polymerase chain reaction kit based assay in comparison to conventional PCR assay and classical microscopy for detection of T. annulata from blood samples of bovines from Punjab state. 
 
Methods: In this comparative study 360 bovine blood samples from various districts of agro-climatic zones in Punjab were first screened for T. annulata by Giemsa-stained thin blood smear (GSTBS) examination. The same panel of blood samples was tested for T. annulata by a commercial PCR kit Bovi-TheiDX Theileria annulata (Genext Genomics) (PCR1) and established conventional PCR assay targeting merozoite piroplasm surface antigen (Tams1) gene of T. annulata (PCR2). 
 
Result:&nbsp;Out of 360 samples screened, positivity of&nbsp;T. annulata by GSTBS was found to be 12.5% (45/360) with a sensitivity of 37.20% and specificity of 96.00% when compared with commercial kit (PCR1), the difference was statistically significant (p&lt;.0001). The detection prevalence by PCR1 29.70% (107/360) and PCR2 assay 33.90% (122/360) showed significant (p&lt;.0001) difference. The conventional PCR targeting the Tams1 gene (PCR2) was found to be more sensitive (84.6%) (p&lt;0.001) than PCR1 (69.7%).</div>

Comparative Performance and Evaluation of Two Molecular Assays and Conventional Detection of Theileria annulata in Bovines

Background: Theileriosis is a tick-borne disease that has a maximum impact on the health and economy of developing countries. A number of clinical manifestations have been found in animals affected with Theileria annulata. A similar trend has been observed in the therapeutic management of theileriosis, especially in cattle. Resistance to available treatment options is one of the significant concerns with theileriosis. 
 
Methods: The present study evaluated the in-vitro activity of moxifloxacin, the fluoroquinolone compound, against T. annulata in 96 well culture plates using resazurin dye. The histopaque method was used for the separation of infected and healthy PBMCs (Polymorph Blood Mononuclear Cells) for growth inhibition assay of T. annulata and cytotoxicity assay measurement, respectively. The hemolytic assay was also evaluated against normal bovine RBCs (red blood cells). The 50% inhibitory concentration (IC50) and 50% cytotoxic concentration (CC50) values were measured and analyzed using a correlation and regression method. 
 
Result: The IC50&nbsp;value of moxifloxacin was observed at 5,559 &micro;M against in-vitro T. annulata. The CC50&nbsp;value of cytotoxicity on PMBCs was found to be 3,984 &micro;M. However, the HC50&nbsp;value of hemolytic assay on bovine red blood cells was observed at 2,59,202 &micro;M. The moxifloxacin showed a wide safety margin on red blood cells. Moxifloxacin can be further investigated in-vivo to determine its actual effect on T. annulata.

In-vitro Activity of Moxifloxacin against Theileria annulata - A Preliminary Study

<div>Background: This study investigated the in vitro activities of naphthoquinones and artemisinin contained in the extracts obtained from Arnebia densiflora and Artemisia annua plants against peripheral blood mononuclear cells infected with Theileria annulata schizonts. For this purpose, the n-hexane extract of A. densiflora and the petroleum ether and methanol extracts of A. annua were obtained. 
 
Methods: Non-infected and infected cells were cultivated and 2&times;106 cells were seeded in each well in 0.5 ml of RPMI 1640 medium. The n-hexane extract of A. densiflora was tested six times at concentrations of 125, 62.5, 15.625, 7.8125 and 3.9062 mg/ml. A. annua petroleum ether and methanol extracts were also tested six times at 125, 62.5, 31.25, 15.625, 7.8125, 3.9062, and 1.9531 mg/ml concentrations on PBMCs. A Mann-Whitney U test was used to determine differences between control and experimental groups. 
 
Result: The A. densiflora extract killed both non-infected and infected cells at significant levels (P&lt;0.05) compared to the control group at all tested concentrations. A. annua methanol extract had no adverse effect on non-infected cells and the effect on infected cells was insufficient, with the only observed significant effect at a concentration of 125 &micro;g/ml. The A. annua petroleum ether extract had no adverse effect on non-infected cells and killed infected cells at significant levels (P&lt;0.05) at concentrations of 15.625 &micro;g/ml and above at 24 hours. At 48 hours, all infected cells were killed at a concentration of 62.5 &micro;g/ml. In conclusion, A. annua petroleum ether extract can be used in future in vivo studies for Theileriosis at these effective concentrations.</div>

In vitro Effect of Arnebia densiflora and Artemisia annua Plant Extracts against&nbsp;Theileria annulata Schizonts

<div>Abortion is the major reproductive problem in goats faced by goat-keepers. Abortion in animals including goats is caused mainly by infectious agents. The current study was planned to assess the presence of Brucella melitensis, Chlamydophila spp., Campylobacter spp., Brucella abortus and Coxiella burnetii in the genital tracts of goats and foetal stomach contents. A total of 260 biological samples (vaginal/preputial secretions and foetal stomach contents) were randomly collected from the goats. The genomic DNA extraction from the samples was done by using the commercial kit. The detection of the targeted infectious agents in the biological samples was done by species as well as genus-specific polymerase chain reactions. Standardization of PCRs was done using gradient PCR. The PCR products were run in a 1% agarose gel to determine the gene size. Out of 260 samples, the presence of Coxiella burnetii was not detected in any sample. Brucella abortus was detected in two aborted goats. Only 10 samples out of 260 (6 vaginal swabs from the aborted goats and 4 foetal stomach contents) were found positive for Brucella melitensis. A total of 117 samples were found positive for Chlamydophila spp., whereas 84 were found positive for Campylobacter spp. A total of 63 samples were found positive for both Chlamydophila spp. and Campylobacter spp. The present study concludes that Chlamydophila and Campylobacter spp. were found as the prevalent microbial agents, which may be associated with abortion in goats in the forthcoming pregnancies in adverse conditions.</div>

Molecular Detection of Brucella melitensis, Chlamydophila spp., Campylobacter spp., Brucella abortus and Coxiella burnetii in Genital Tracts of Goats

<div style="text-align:justify">Background: The Pleurotus genus is one of the most investigated white-rot fungus due to its excellent ligninolytic characteristics. It is a tasty mushroom that also has a number of biological impacts due to the presence of essential bioactive compounds. Many usual fermentation parameters affect lignocellulolytic enzymes in basidiomycete fungi, such as medium composition, carbon-to-nitrogen ratio, pH, temperature, air composition etc. The survival and multiplication of Pleurotus mushrooms is dependent on a number of factors, which may function independently or interactive effect on the growth of mushroom. Grasp the difficulty of treating Pleurotus species mushrooms necessitates a fundamental understanding of their physical, chemical and biological characteristics.</div>

<div style="text-align:justify">Methods:&nbsp; In this study, the effects of culture media, temperature, carbon sources, nitrogen sources, grain sources and agricultural waste as lignocellulosic substrate on the mycelium growth of five species of Pleurotus species for spawn production were studied. Several trials were designed to evaluate factors affecting mycelium growth of five oyster mushroom species. The experiments were conducted during October to February in the year 2019 to 2021. The experiments were carried out in three replications per treatment. 
Result: The results of the experiment indicated that potato dextrose agar (PDA) and Malt extract agar (MEA) were the most suitable media for the mycelium growth of all oyster mushrooms. The optimal temperature for mycelium growth of all the oyster mushroom species was obtained at 25&plusmn;2oC. Mycelium growth of all the studied oyster mushroom was improved by carbon sources such as dextrose and glucose. The nitrogen source such as ammonium chloride and ammonium sulfate also gave the greatest values in mycelium colony diameter of mushroom. Wheat was found to be favourable for mycelium growth of all the oyster mushroom species. In addition, paddy straw and sugarcane residue were selected as suitable lignocellulosic substrate sources for mycelium growth of all the studied oyster mushrooms.</div>

The Effect of Growth Conditions on Mycelial Run of Oyster Mushrooms spp. (Pleurotus spp.): Implication for Agricultural Practices

<div>Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India.&nbsp; 
 
Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman&rsquo;s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis. 
 
Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.</div>

Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

<div style="text-align:justify">Background: Theileriosis is a haemoprotozoan illness that poses severe economic loss to dairy farmers due to its negative impact on mortality and productivity, especially in tropical and subtropical areas of the world. 
 
Methods:&nbsp;The present study was conducted for a period of one year (June 2020 to July 2021) during which 55 blood samples of cattle showing clinical signs suggestive of theileriosis were examined and quantified by using multiplex real time PCR for Theileria orientalis and Theileria annulata using Taq-Man probe. 
 
Result: Prevalence of theileriosis in three Theileria-endemic districts of Odisha due to Theileria annulata and Theileria orientalis were 26.11 per cent (11/42) and 57.14 per cent (24/42), respectively. Mixed infection were reported in 16.6 percent (7/42) cases. Parasite load for T. annulata in clinically affected cases ranged from 2.85&times;103 to 1.51&times;105 parasites per ml of blood while for T. orientalis,&nbsp;it ranged from 1.94&times;104&nbsp;to 5.88&times;105&nbsp;parasites per ml of blood. Ten cattle positive for T. orientalis were administered two doses of buparvaquone (Butalex) @2.5 mg/kg b.w. at an interval of 48 hours after which degree of parasitemia were quantified ten days post treatment. The parasitic load in blood ten days post treatment decreased significantly from 4.04&times;104 to 2.21&times;102 per ml of blood.</div>

RT PCR Assay to Determine Efficacy of Buparvaquone in Reducing Degree of Parasitemia of Theileria orientalis

<div>Background: China is a mainland country rich in natural Morchella spp. resources. At present, about half of the natural Morchella spp. in the world has been recorded in China. The current study was aimed at the classification and cultivation of Morchella spp. This study provided a more accurate molecular trait for the systematic classification of Morchella spp. in southern Gansu Province, China. 
 
Methods: From April to May 2019. Based on the molecular biological analysis of 16 natural Morchella spp. strains collected from the southern Gansu province, China. the ITS fragments between transcripts of the rDNA gene were amplified by PCR and sequenced. 
 
Result: The results of BLAST comparison according to GenBank showed that the 16 tested strains of Morchella spp. were classified into 5 species, which were M.angusticeps, M.esculenta, M.elata, M.crapssipes&nbsp;and M.conica. According to the molecular evolutionary trees constructed by the Maximum-Parsimony method (MP) and the Neighbour-Joining method (NJ), the topological structures of the two molecular evolutionary trees were similar&nbsp;and the Bootstrap verification had a high support rate, indicating that the phylogenetic relationship had high credibility.</div>

<div>Genetic Diversity of Natural Morchella spp. from the Southern Gansu Province Based on rDNA-ITS, China</div>

<div style="text-align: justify;">Chickpea (Cicer arietinum L.) is the third most important legumes after beans and peas in the world. Turkey is one of the main chickpea producing countries but the yield is not within the expected quantity considering cultivated area. Breeding chickpea varieties requires well-characterized wild Cicer spp progenitors including C. reticulatum, C. echinospermum, C. bijugum and C. pinnatifidum that exhibit sympatric distribution in the Southeastern Anatolia region of Turkey. This study contributes to the identification of similarities and differences among wild and cultivated Cicer spp. ecotypes using DNA marker systems and biochemical parameters. RAPD and ISSR markers revealed four groups and separated C pinnatifidum ecotypes as a single population. Wild Cicer spp and cultivated chickpea species exhibited significant variations for their oil, protein, starch, cellulose, fructose and sucrose content. Furthermore, the cultivated chickpea genotypes had higher content of fat, starch and sucrose than those of the wild progenitors whereas wild Cicer spp ecotypes possessed higher values for cellulose, fructose and glucose than cultivated chickpea genotypes.&nbsp;</div>