Furkan Alaraji, Hussam Muhsen, Abdullah O. Alhatami and Yahia Ismail Khudhair
For the first time in Iraq, we identified in March, 2018 the presence of a highly virulent avian influenza virus (AIV), H5N1 (Clade 18.104.22.168c), causing highly pathogenic avian influenza (HPAI) in poultry farms, Iraq,. The identification of the virus was done using a rapid serological test, a real time-qPCR, and glycoprotein gene sequencing. Using sequencing and phylogenetic analyses, the clade 22.214.171.124c virus was recorded to be clustered, with high similarity to Asian and West African AIV, HPAI H5N1 from Ivory Coast identified in 2015. According to our knowledge, there was no previous detection of the clade 126.96.36.199c made in Iraq. Our results provide evidence that high risk of HPAI H5 outbreaks might be present in Iraq, and this needs to lead to high quality surveillance targeting of wild and domestic birds for early diagnosis of HPAI. The current work provides feasible and accurate approaches for understanding the evolution of HPAI H5 virus in different countries around the world.
Hayder N. Ayyez, Yahia I. Khudhair and Qassim Haleem Kshash
Anaplasma spp. are widely spread rickettsial bacteria transmitted by ticks and placing high impacts on veterinary and public health. A limited number of studies have been carried out on Anaplasmosis in the central part of Iraq. This study was conducted to determine the presence of Anaplasma spp. in cattle in Al-Qadisiyah province, Iraq. A total of 400 blood specimens were collected from cattle suffering from heavy tick infestation. Cattle were blood-sampled from four hyperendemic areas with ticks. Blood samples were screened using microscopic and polymerase chain reaction (PCR) methods. Diff-quick stained blood smears revealed Anaplasma-like inclusion bodies in 254 (63.5%) samples. According to the 16S rRNA-gene-based PCR analysis, Anaplasma spp. was detected in 124 of the 400 (31%) samples, divided as 96/254 (37.8%) among the microscopical positive samples and 28/146 (19.17%) among the microscopical negative samples. Phylogenetic analysis based on the partial 16S rRNA gene sequencing of ten-PCR positive samples were 99–97% identical to sequences deposited in the GenBank, revealing presence of A. phagocytophilum, A. marginale and unnamed Anaplasma spp. in 40%, 20%, and 40% samples, respectively. Relationships among Anaplasma spp. infections and cattle breed, age, and sex were analyzed. Calves less than one year old showed significantly higher rates (p<0.005) than those from other age groups, whereas sex and breed demonstrated no significant differences (p˃0.001). This study shows that a variety of Anaplasma spp., were endemic in central part of Iraq and is still a hidden problem in cattle in the hyperendemic areas of tick, which requires serious control strategies.
Martin Nikolovski, Monika Dovenska, Ksenija Ilievska, Nikola Adamov, Branko Atanasov, Miroslav Radeski, Daniela Kirovski, Vladimir Petkov and Toni Dovenski
Reduced glutathione (GSH) and homologous ram seminal plasma (HSP), used as additives in cryopreserving (CP) media prior to freezing, showed conflicting results in retaining structural integrity and progressive motility in post-thawed ram spermatozoa. The aims of this research were: (1) to assess the effect of GSH and/or HSP supplementation via soybean-lecithin CP extender on cryopreserved ram spermatozoa viability, morphology and motility pattern; and (2) to assess the effect of incubation in the context of the previous aim. Quantitatively and qualitatively, homogenized and pooled ram ejaculates (N=10) were extended with one of the following extenders: control (C) – tris-based, GSH and HSP-free, experimental-1 (E1) – C + GSH 5 mM, experimental-2 (E2) – C + HSP 20 % and experimental-3 (E3) - GSH 5 mM + HSP 20 %. Following thawing, samples were taken at 0- and 3-hours from each group (n=10) and were assessed for spermatozoa viability, morphology, and motility pattern. C-0h samples yielded a spermatozoa population with low viability, altered head morphology and highly deviated motility pattern. E3-3h samples yielded spermatozoa with unaffected viability, head morphology and high progressive motility. In conclusion, E3 extender added to cryopreserved-thawed ram spermatozoa is most efficient in obtaining high viability, unaltered head morphology, and progressive motility.
Snežana Paskaš, Jelena Miočinović, Mila Savić, Goran Ješić, Mladen Rašeta and Zsolt Becskei
This study was conducted to evaluate the nutritional quality of two types of cheese prepared from bovine whey: Urda and Ricotta. Cheeses were examined for chemical parameters and their amino acids profile was determined. The results revealed that the chemical composition of the observed whey cheeses was highly significantly different (p<0.01) from each other. The amounts of dry matter, fat, protein, lactose, ash and salt were higher in Ricotta compared with Urda cheese samples. On the other hand, Urda contained more moisture, fat on a dry matter basis and moisture on a fat-free basis (79.59%, 27.50%, 84.27% versus 69.82%, 21.02%, 74.56%, respectively). Especially higher yield, salt in moisture content and energy values were observed in Ricotta cheese (5.93%, 3.12%, 145.99Kcal/100g versus 4.39%, 2.40%, 108.97Kcal/100g; Ricotta and Urda, respectively). Whey cheeses are a particularly good source of amino acids containing approximately - Ricotta: leucine (1.60g/100g), lysine (1.17g/100g), phenylalanine (0.78g/100g) and followed by threonine (0.77g/100g), while Urda contained leucine (0.80g/100g), lysine (0.45g/100g), threonine (0.68g/100g) and phenylalanine (0.31g/100g). Both cheese varieties contained glutamic and aspartic acid as the predominant non-essential amino acids (Ricotta: 3.91g/100g and 1.68g/100g versus Urda: 1.65g/100g and 0.80g/100g, respectively). Generally, from a nutritional point of view, the investigated whey cheeses could be considered as cheeses with low salt and fat content, as well as cheeses particularly abundant in branched chain amino acids (leucine, isoleucine and valine). These results also emphasize their advantages as products that present a cost-effective way of dealing with whey as waste material.
The vascular endothelial growth factor (VEGF) is a multifunctional cytokine stimulating the growth of vascular endothelial cells, survival and proliferation, inhibiting apoptosis. It is one of the most potent stimulants of vascular permeability. VEGF is found at high levels in inflammatory and tumour-associated pleural and abdominal effusions and is involved in their occurrence. In the present study, the blood plasma and abdominal fluid VEGF levels were assayed in thirty-one client-owned dogs with neoplastic and non-neoplastic diseases by means of enzyme-linked immunosorbent assay (ELISA). The VEGF concentration in abdominal fluid of dogs (n=6) with ascites was 190.70±34.35 pg/ml, in dogs (n=6) with peritonitis: 1449.81±365.42 pg/ml and in dogs (n=9) with tumour-associated effusion: 1993.13±202.56 pg/ml. Blood plasma VEGF of healthy dogs (control group, n=10) was 36.79±5.72 pg/ml, in dogs with ascites: 57.92±2.88 pg/ml, in dogs with peritonitis: 76.98±7.24 pg/ml and in dogs with tumour-associated effusion: 173.50±40.9 pg/ml. There were substantial differences between blood plasma and abdominal fluid VEGF levels.
A 15 year old female African lion (Panthera leo) was necropsied after its sudden death. The necropsy showed a mammary gland lesion measuring 10 cm in diameter and numerous white nodules with variable size in the liver, spleen, uterus, lungs and the heart. The histopathological examination showed that the neoplastic formation in the mammary region was a simple tubular carcinoma with metastases on the other organs. Upon immunohistochemical examination, the neoplastic cells expressed cytokeratins while the stroma of the tumour expressed vimentin. The proliferation index Ki-67 was moderate. Based on the macroscopic, histopathological and immunohistochemical findings, the neoplasia was diagnosed as a simple tubular mammary carcinoma.
Alhaji Zubair Jaji, Adamu Saleh Saidu, Mohammed Bakari Mahre, Mbaya Pindar Yawulda, Ibrahim Alhaji Girgiri, Piyush Tomar and Faruk Dau
Prenatal gross morphologic, morphometric and histologic developmental features of the dromedary spleen were studied. The dromedary gestation period (13 months) was categorized into four (1-4) phases and ten developing spleens per growth phase were sampled. Splenic topographical anatomy was noted before being eviscerated from each foetus. Morphologic and morphometric features of the eviscerated spleens were immediately documented and 2 – 4 mm thick samples were collected for histological analysis. The developing spleen was dark brown in colour, semilunar shaped and significantly increased (p<0.05) in size and weight across the four phases of prenatal development. The full-term dromedary spleen was observed to have unique histological features. Its capsule had an inner smooth muscle and an outer predominant connective tissue layer. The pumping of stored blood from the muscular capsule and trabeculae was proportionate to the body’s requirement. The splenic venous return was characterized by blood flow from the red pulp (venous sinusoids) to the peritrabecular sinuses, subcapsular sinuses and finally to the splenic veins. The dromedary has a sinusal type of spleen and has both open and closed types of circulation. The presence of closed circulation and absence of marginal sinus could be the reason for dromedary main health problems of blood parasites; Trypanosoma evansi. It was concluded that most of the salient features of the postnatal spleen were already evident in the first growth phase and became developed by the second phase. Other growth phases were mainly characterized by increase in sizes.
A fast and sensitive HPLC method was validated in order to analyze doxycycline in plasma and milk of sheep and in plasma of rabbits. The samples were processed with trifluoroacetic acid (TFA). After the centrifugation step, a supernatant containing extracted doxycycline and internal standard oxytetracycline was injected into the HPLC system with PDA detection. The method showed linearity in the range of 0.125 - 2.5 μg/mL for ovine plasma, 0.125 – 5.0 μg/mL for ovine milk, and 0.125 – 1 μg/mL for rabbit plasma. The inter-assay precision varied between 5.69 – 13.55 %. Values for intraassay precision were between 0.62 – 8.67 %. Accuracy was higher than 90% in all of the tested concentrations in the three types of biological matrices. The mean extraction recovery was higher than 90 % for all matrices. In order to handle only with free drug concentrations, microfiltration of standard solutions with low (0.25mg/mL), medium (0.5mg/mL) and high (1.0mg/mL) concentration was performed. A percentage for correction of the quantified doxycycline was calculated. The most significant adjustments should be made at the low concentrations. The correction for rabbit plasma is 24.63±5.03%, for ovine plasma is 20.10±8.01% and for milk –16.68±0.04 %. This method can be used for routine determination of doxycycline concentrations for pharmacokinetic studies and further dosage adjustment.
Ivaylo Sirakov, Katya Velichkova, Stefka Stoyanova, Georgi Zhelyazkov and Yordan Staykov
The aim of the current study was to test the effect of a diet supplemented with dandelion’s (Taraxacum officinale) extract on the productive traits and blood parameters in common carp (Cyprinus carpio L.) cultivated in an recirculation aquaculture system. The carps were cultivated at a stocking density of 7.2 kg/m3 in recirculation aquaculture system. The fish were split into the following two experimental groups: DF (the fish were fed with feed supplemented with dandelion’s extract) and CF (the fish were fed with feed without supplementation). Common carp fed with a diet added with dandelion’s extract at a quantity of 0.8% from daily feed ratio, did not affect the hydrochemical parameters (pH, dissolved oxygen, and electrical conductivity). The carps fed with feed supplemented with dandelion’s extract did not show better productive traits compared with these found for carp from the control variant. The carp from experimental groups had a higher survival rate, final weight, average individual weight gain and specific growth rate (SGR), respectively with 13.2%, 3.94%, 31.5% and 31.3%, compared with the average values of these parameters measured in individuals fed with the control feed, but the differences were not statistically significant (P≥0.05). Supplementation of feed with dandelion’s extract significantly decreased the plasma cholesterol (4.76%) and triglyceride (61.2%) content, promoting hypolipidemic status in fish (P≤0.05).
Aleksandar Dodovski, Zagorka Popova and Vladimir Savić
Avian avulavirus type 1 (AAvV-1) belongs to the family Paramyxoviridae, genus Avulavirus. Even though belonging to a single serotype, there is great genetic variability between these viruses. On the basis of the available complete fusion (F) gene and whole genome sequences and using the genotype classification system, AAvV-1 viruses are divided in two major groups: class I and class II. Class I viruses are predominantly avirulent viruses and majority of the isolations come from waterfowl and domestic poultry from live bird markets in USA and China although isolations from wild birds are reported globally. In our study we used classical, molecular and phylogenetic tools to characterize an AAvV-1 isolated from a mute swan in Macedonia. Based on the complete F gene sequence, we have concluded that the virus designated as AAvV-1/mute swan/Macedonia/546/2012 (KP123431) belongs to the class I of AAvV-1 with an avirulent cleavage site motif 112ERQER*L117. The virus could not be assigned to any of the four currently existing subgenotypes (1a, 1b, 1c or 1d) of the single genotype 1 of class I viruses. Two distant viruses, isolated from goose in Alaska in 1991 and from goose in Ohio in 1987, shared the highest nucleotide sequence identity of the complete F gene with the isolate in our study: 92.7% and 92.8%, respectively. This is the first report of isolation of class I AAvV-1 in Southeastern Europe. The asymptomatic nature and the underreporting of sequences from the class I viruses impede the understanding of the molecular epidemiology and evolution of this group of viruses.