Ficus sycomorus latex: An efficient alternative Egyptian source for horseradish peroxidase in labeling with antibodies for immunodiagnostic kits

Research (Published online: 01-10-2018)

1. Ficus sycomorus latex: An efficient alternative Egyptian source for horseradish peroxidase in labeling with antibodies for immunodiagnostic kits

Azza M. Abdel-Aty, Mohamed Belal Hamed, Abdul Aziz M. Gad, Amr E. El-Hakim and Saleh A. Mohamed

Veterinary World, 11(10): 1364-1370

ABSTRACT

Aim: In view of various peroxidase applications, the searching for new sources of unique peroxidase properties is highly desirable. The present study aims to evaluate the efficiency of the peroxidase of locally grown sycamore latex (POL) for conjugation with antibodies and to study the conjugate optimal conditions, storage stability, and affinity toward different substrates as compared with commercial horseradish peroxidase (HRP).

Materials and Methods: Anti-mouse antibodies were prepared in rabbits and purified by protein A sepharose affinity column chromatography. The POL and HRP conjugates were prepared by one-step glutaraldehyde coupling method. The reactivity of the prepared conjugates was examined using the enzyme-linked immunosorbent assay (ELISA). The optimal enzymatic conditions, storage stability, and affinity toward substrates were also determined for both the conjugates.

Results: The POL showed higher percent recovery (98%) than HRP (78%) over the initial activity after conjugation process. The POL and HRP conjugates showed ELISA titers of 1:120 and 1:80, respectively, demonstrating high binding affinity of POL-conjugate. The POL-conjugate showed high thermal stability up to 70°C compared with HRP-conjugate up to 40°C. After conjugation, POL had wide pH stability (5.0-8.0) compared with HPR (4.5-6.0). Both of the prepared conjugates had a high affinity toward the substrates used in immunoassays with lower Km values. The POL-conjugate showed high storage stability for its enzymatic activity and ELISA titer compared with HRP-conjugate after 1 year at −20°C.

Conclusion: The POL of Ficus sycomorus latex is an efficient source for labeling antibodies and could be utilized in immunodiagnostic kits.

Keywords: conjugation, Ficus sycomorus, horseradish peroxidase, immunodiagnostic assays, latex.

Hard tick (Acari: Ixodidae) species of livestock and their seasonal activity in Boyer-Ahmad and Dena cities of Kohgiluyeh and Boyer-Ahmad Province, Southwest of Iran

Research (Published online: 30-09-2018)

25. Hard tick (Acari: Ixodidae) species of livestock and their seasonal activity in Boyer-Ahmad and Dena cities of Kohgiluyeh and Boyer-Ahmad Province, Southwest of Iran

Zohreh Fatemian, Aref Salehzadeh, Mohammad Mehdi Sedaghat, Zakieh Telmadarraiy, Ahmad Ali Hanafi-Bojd and Amir Hosein Zahirnia

Veterinary World, 11(9): 1357-1363

ABSTRACT

Aim: The present study was carried out to identify the Ixodidae ticks fauna of livestock and their seasonal activity in the cities of Boyer-Ahmad and Dena of Kohgiluyeh Province, south-west of Iran.

Materials and Methods: Hard ticks from sheep, goats, and cattle were collected manually, stored in 70% ethanol, and identified using morphological characters.

Results: During the study, a total of 1273 hard ticks from four genera, including Rhipicephalus, Hyalomma, Dermacentor, and Haemaphysalis, were collected. Rhipicephalus sanguineus (s.l.) had the highest frequencies in both cities with 62.08 and 62.88% of collected specimens, followed by Hyalomma scupense with 14.36 and 13.54% in Boyer-Ahmad and Dena, respectively. Furthermore, Hyalomma marginatum with only one sample or 0.12% of collected ticks showed the lowest frequencies in the studied areas. Dermacentor marginatus with three samples or 0.37% was recorded only in Boyer-Ahmad, and Haemaphysalis sulcata with two samples or 0.43% was recorded only in Dena. In both cities, sheep were the most infested ruminant, and the ears in sheep and goats were the most affected areas. The highest activity was observed in spring, and the lowest activity was observed in winter and autumn.

Conclusion: The results of the present study showed that Hyalomma and Rhipicephalus genera were the most widespread genera in the study areas. Regarding the importance of genera, such as Rhipicephalus, Hyalomma, and Haemaphysalis, in transmitting disease agents and the location of Kohgiloyeh and Boyer-Ahmad Province in the routes of migrant birds, further studies are required to elucidate their exact roles in human and livestock health in these areas.

Keywords: domestic ruminants, fauna, Ixodidae ticks, seasonal activity, tick-borne diseases.

Effects of secretome on cisplatin-induced testicular dysfunction in rats

Research (Published online: 29-09-2018)

24. Effects of secretome on cisplatin-induced testicular dysfunction in rats

Surya Agus Prihatno, Irma Padeta, Arinda Devi Larasati, Betty Sundari, Annisa Hidayati, Yuda Heru Fibrianto and Teguh Budipitojo

Veterinary World, 11(9): 1349-1356

ABSTRACT

Background: Testicular dysfunction is a degenerative disorder characterized by failure in the synthesis of reproductive hormones and spermatogenesis. Secretome derived from the human umbilical mesenchymal stem cell (MSC) has been reported to repair some degenerative disorders.

Aim: This study aimed to investigate the effect of secretome derived from the human umbilical MSCs on cisplatin-induced testicular dysfunction in rats.

Materials and Methods: Thirty-six male Wistar rats were divided into the control and secretome-treated groups. In the secretome-treated group, testicular dysfunction was induced by 3 mg/kg BW of cisplatin intraperitoneally 3 times with 3-day intervals. The secretome-treated group was divided according to dose: Low-dose (0.2 mL/kg BW) and high-dose (0.5 mL/kg BW) groups. Secretomes were injected intraperitoneally once a week for 3 weeks. 1 week after the injection of secretome, the cauda epididymis of the rats was removed for spermatozoa evaluation and histological examination.

Results: After the injection of secretome, the sperm motility of the high-dose group showed thin wave-like, rare, and slow movements. No abnormal sperm morphology was observed in all the treated groups. The number of spermatozoa increased gradually in the high-dose group after the injection of secretome. The developmental stages of the spermatogenic cells were complete in both spermatozoa groups after the injection of secretome. However, the spermatozoa in the seminiferous tubules of the high-dose group were denser. Vimentin and cytokeratin immunoreactivities were very strong in the high-dose group 1 week after the second secretome injection.

Conclusion: High-dose secretome derived from the human fetal umbilical cord could increase the number and motility of sperms in rats with cisplatin-induced testicular dysfunction. The administration of high-dose secretome was effective 1 week after the second dose, as indicated by very strong immunoreactivity for vimentin and cytokeratin. Moreover, secretome could promote the regeneration of the seminiferous tubules of both the groups.

Keywords: cisplatin, cytokeratin, secretome, spermatogenesis, testicular dysfunction, vimentin.

Dietary encapsulated probiotic effect on broiler serum biochemical parameters

Research (Published online: 29-09-2018)

23. Dietary encapsulated probiotic effect on broiler serum biochemical parameters

P. Yazhini, P. Visha, P. Selvaraj, P. Vasanthakumar and V. Chandran

Veterinary World, 11(9): 1344-1348

ABSTRACT

Aim: The study aimed to evaluate the effect of encapsulated probiotic bacteria (Lactobacillus lactis and Bifidobacterium bifidum) on broiler serum biochemical parameters.

Materials and Methods: Encapsulation protects the probiotics and increases their livability on exposure to unfavorable processing and storage temperatures and gastrointestinal pH. Hence, an in vitro study was undertaken to encapsulate the probiotic bacteria L. lactis and B. bifidum with sodium alginate and chitosan and evaluate the encapsulation efficiency. This experiment was conducted with 288-day-old broiler chicken; they were distributed randomly into eight treatments and six replicates in each treatment (six birds in each replicate) and given with standard feed.

Results: Supplementation of the encapsulated bacteria either alone or in combination (T4, T6, and T8) significantly (p<0.05) increased mean total serum protein, albumin, and globulin as compared to the birds that were not supplemented with any probiotic (T1 and T2) or supplemented with non-encapsulated bacteria (T3, T5, and T7). Supplementation of the encapsulated bacteria either alone or in combination (T4, T6, and T8) significantly (p<0.05) lowered mean total serum cholesterol, serum low-density lipoprotein (LDL) cholesterol, and serum triglycerides, as compared to the birds that were not supplemented with any probiotic (T1 and T2) or supplemented with non-encapsulated bacteria (T3, T5, and T7).

Conclusion: It may be concluded that supplementation of the encapsulated probiotic bacteria either alone or in combination significantly increased total serum protein, albumin, and globulin and significantly lowered mean total serum cholesterol, serum LDL cholesterol, and serum triglycerides as compared to the birds that were not supplemented with any probiotic or supplemented with non-encapsulated bacteria.

Keywords: biochemical, broiler, encapsulated, probiotic, serum.

Detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction using repetitive 529 bp gene in feces of pet cats (Felis catus) in Yogyakarta, Indonesia

Research (Published online: 28-09-2018)

22. Detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction using repetitive 529 bp gene in feces of pet cats (Felis catus) in Yogyakarta, Indonesia

Muhammad Hanafiah, Joko Prastowo, Sri Hartati, Dwinna Aliza and Raden Wisnu Nurcahyo

Veterinary World, 11(9): 1338-1343

ABSTRACT

Aim: The aim of this research was to determine the copro-prevalence of Toxoplasma gondii using polymerase chain reaction (PCR) with repetitive 529 bp gene and to construct the phylogenetic tree of Toxoplasmaoocyst from pet cats in Yogyakarta.

Materials and Methods: 9 of 132 pet cat samples which serologically positive for Toxoplasma were used in this research. To determine the copro-prevalence of T. gondii in pet cat, 10 g of feces samples taken from practitioners and household cats in Yogyakarta were used in the PCR method utilizing repetitive 529 bp gene sequences.

Results: The result shows that copro-prevalence by PCR using repetitive 529 bp gene was 33.3% (3/9). The phylogenetic tree of Toxoplasma grouped into two clades, which clade 1 consists of Toxoplasma isolates collected from pet cats in Yogyakarta Indonesia and T. gondii isolates from China and in clade 2 consist of the T. gondii isolates from India.

Conclusion: Copro-prevalence of T. gondii in pet cats in Yogyakarta by means of PCR using repetitive 529 bp gene is around 33.3%.

Keywords: copro-prevalence, pet cat, polymerase chain reaction, Toxoplasma gondii.

Epidemiological surveillance of bovine viral diarrhea and rift valley fever infections in camel

Research (Published online: 26-09-2018)

21. Epidemiological surveillance of bovine viral diarrhea and rift valley fever infections in camel

Halla E. K. El Bahgy, Hala K. Abdelmegeed and Marawan A. Marawan

Veterinary World, 11(9): 1331-1337

ABSTRACT

Aim: This study was designed to investigate the current epidemiological situation of bovine viral diarrhea virus (BVDV) and rift valley fever virus (RVFV) infection of camels originating from Sudan "smuggler" and Egypt as part of our future plan for a national surveillance program in Egyptian provinces, which will aid in establishment of control strategy for animal diseases.

Materials and Methods: This investigation was accomplished using serological diagnostic and molecular biology techniques. A total number of 200 blood samples were collected from camel (120 originated from Sudan "smuggler" and 80 from local breed) and were subjected for testing both BVDV and RVFV occurrence with different age and sex.

Results: Sixty-six of the 200 camels (33%) were positive for BVDV antibodies, and 44 (22%) for BVDV antigen (Ag), and 27 of the 200 camels (13.5%) were positive for RVFV immunoglobulin G (IgG) antibodies. On the other hand, the seroprevalence of BVDV for antibodies (47.5%), Ag (31.6%), and RVFV IgG antibodies (16.6%) was higher in camel originated from Sudan "smuggler" than of local breed which was 11.2% for BVDV antibodies and 7.5% for BVDV Ag, while it was 8.7% for RVFV IgG antibodies. The incidence of BVDV antibodies, Ag, and RVFV IgG antibodies was the highest in male, up to 9 years of age. The frequency of positive cases was significantly different according to the origin of samples and sex and age of camel for BVDV and RVFV. In addition, seven serologically positive samples for BVDV and five serologically positive samples for RVFV were submitted as a buffy coat for molecular detection by one-step - reverse transcription polymerase chain reaction (RT-PCR). The results demonstrated that three samples were positive for BVDV of camel originated from Sudan (smuggler), while no RVFV Ag was detected in all five samples. Sequencing and phylogenetic analysis of the amplicons obtained from positive RT-PCR samples (three samples) indicated 100% nucleotide homology with Sudan strain 2015 except only one (missense point mutation) by substitution of A to T at position 345 that changed the coded amino acids from T (Threonine) to S (Serine) at residue 115.

Conclusion: Camels act as risk animals for the introduction of many infectious diseases from Sudan to Egypt, especially transboundary animal diseases, so strict quarantine measures should be taken during importation of live animals from Sudan to prevent the spread of such diseases.

Keywords: bovine viral diarrhea virus, epidemiological, rift valley fever virus, risk factors, sequencing, surveillance.

First-time serological and molecular detection of Helicobacter pylori in milk from Algerian local-breed cows

Research (Published online: 25-09-2018)

20. First-time serological and molecular detection of Helicobacter pylori in milk from Algerian local-breed cows

Meryem Guessoum, Zehor Guechi and Mounir Adnane

Veterinary World, 11(9): 1326-1330

ABSTRACT

Aim: The present study was conducted to detect and identify Helicobacter pylori within local cow breeds in the central region of Algeria.

Materials and Methods: Two hundred (n=200) cows from three provinces of the central region of Algeria were studied, between January 2016 and September 2017. Each cow was subject to stool, milk, and blood sampling. Milk and fecal samples were used to detect and identify H. pylori using bacteriology culture method. Blood and milk samples were used to detect H. pylori immunoglobulin G (IgG) antibody using enzyme-linked immunosorbent assay. Polymerase chain reaction was used to confirm the abundance of H. pylori in milk by detecting glmM gene.

Results: Out of 200 sera and 200 milk samples, 12% (24) and 4% (8/200) were positive for the H. pylori IgG antibody. glmM gene was detected in the milk of 13% of cows and was confirmed in all cows presenting IgG in milk.

Conclusion: From the present study, we concluded that the glmM gene is an important marker for detecting H. pylori in milk. Moreover, Algerian local-breed cows are a source of H. pylori and could be responsible for serious zoonosis.

Keywords: Algeria, cows, feces, Helicobacter pylori, milk, serum.