Triple gene editing in porcine embryos using electroporation alone or in combination with microinjection

Research (Published online: 27-02-2022)

33. Triple gene editing in porcine embryos using electroporation alone or in combination with microinjection

Zhao Namula, Quynh Anh Le, Manita Wittayarat, Qingyi Lin, Koki Takebayashi, Maki Hirata, Lanh Thi Kim Do, Fuminori Tanihara and Takeshige Otoi

Veterinary World, 15(2): 496-501

ABSTRACT

Background and Aim: We previously developed the gene-editing by electroporation (EP) of Cas9 protein method, in which the CRISPR/Cas9 system was introduced into porcine in vitro fertilized (IVF) zygotes through EP to disrupt a target gene. This method should be further developed, and a combination of EP and MI methods should be evaluated in pigs. This study aimed to determine that a combination of microinjection (MI) and EP of CRISPR/Cas9 system could increase the rates of biallelic mutation for triple-gene knockout in porcine blastocysts. We targeted the pancreatic and duodenal homeobox1 (PDX1) gene using cytoplasmic MI 1 h before or after EP, which was used to edit alpha-1,3-galactosyltransferase (GGTA1) and cytidine 32 monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes in porcine zygotes.

Materials and Methods: We introduced guide RNAs targeting PDX1, GGTA1, and CMAH with the Cas9 protein into IVF zygotes (one-cell stage) through EP 10 h after the start of IVF (IVF; EP group) or in combination with MI (1 h before, MI-EP group, or after EP treatment EP-MI group) and evaluated the blastocyst formation rate and efficiency of target mutations in the resulting blastocysts.

Results: Our results revealed a significant reduction in the rate of blastocyst formation in the two groups that underwent MI before and after EP (MI-EP and EP-MI group), compared with that in the groups treated with EP alone (EP group) (p=0.0224 and p<0.0001, respectively) and control (p=0.0029 and p<0.0001, respectively). There was no significant difference in the total mutation rates among the treatment groups in the resulting blastocysts. As an only positive effect of additional MI treatment, the rate of blastocysts carrying biallelic mutations in at least one target gene was higher in the MI-EP group than in the EP group. However, there was no difference in the rates of embryos carrying biallelic mutations in more than 2 target genes.

Conclusion: These results indicate that although a combination of MI and EP does not improve the mutation efficiency or biallelic mutation for triple-gene knockout, MI treatment before EP is better to reduce mortality in porcine zygotic gene editing through a combination of MI and EP.

Keywords: clustered regularly interspaced short palindromic repeats/Cas9, electroporation, gene editing, microinjection, porcine zygotes.

Development of multidrug-resistant Escherichia coli in some Egyptian veterinary farms

Research (Published online: 27-02-2022)

32. Development of multidrug-resistant Escherichia coli in some Egyptian veterinary farms

A. A. Samy, Asmaa S. Mansour, Doaa D. Khalaf and Eman A. Khairy

Veterinary World, 15(2): 488-495

ABSTRACT

Background and Aim: Food of animal origin is considered a major source of foodborne diseases. In this context, multidrug-resistant (MDR) Escherichia coli pose a serious hazard to public health due to the consumption of food contaminated with antibiotics that are used for the treatment of various bacterial infections in farm animals. Therefore, this study aimed to determine the effect of the excessive use of antibiotics on the development of MDR E. coli strains in Egyptian poultry, dairy, and meat farms.

Materials and Methods: A total of 1225 samples were randomly collected from poultry, dairy, and meat products intended for human consumption in different governorates. E. coli were isolated from the collected samples and subjected to biochemical identification and antibiotic sensitivity tests with antibiotics commonly used in human and veterinary medicine. Then, amoxicillin (AML)- and oxytetracycline (OT)-resistant E. coli isolates were subjected to a polymerase chain reaction test to detect the blaTEM and tetA genes, respectively.

Results: E. coli were isolated from 132 out of 350, 148 out of 350, 177 out of 350, and 35 out of 175 poultry, milk, meat, and human samples, respectively. Most of the isolates expressed multidrug resistance, and resistance genes (blaTEM and tetA) were detected in all the tested AML- and OT-resistant E. coli isolates.

Conclusion: Foods of animal origin may represent a source of MDR E. coli, which can be a major threat to public health.

Keywords: Escherichia coli, food of animal origin, misuse of antibiotics, multidrug resistance, poultry.

Canine testicular tumors: An 11-year retrospective study of 358 cases in Moscow Region, Russia

Research (Published online: 26-02-2022)

31. Canine testicular tumors: An 11-year retrospective study of 358 cases in Moscow Region, Russia

Aleksey A. Gazin, Yury A. Vatnikov, Nikolay V. Sturov, Evgeny V. Kulikov, Viktor Grishin, Elena A. Krotova, Alisa A. Razumova (Varentsova), Natalia Yu. Rodionova (Sapego), Natalia I. Troshina, Varvara M. Byakhova and Ksenia V. Lisitskaya

Veterinary World, 15(2): 483-487

ABSTRACT

Background and Aim: Canine testicular tumors are among the most common reproductive tract tumors in male dogs and have been studied in many countries. However, to the best of our knowledge, studies with a large sample size have not been conducted in Russia. This study aimed to provide the latest information on the prevalence of canine testicular tumors in the Veterinary Oncology Scientific Center for Small Animals "Biocontrol" in Moscow, Russia, in 2010-2020 and the characteristics of the affected canine population.

Materials and Methods: A retrospective review of patients and histological reports was collected and analyzed from 358 dogs with 447 testicular tumors within 11 years.

Results: The mean age of the affected dogs was 10.4 years, whereas that of dogs with Sertoli cell tumors was 9.4 years p=0.009. This study includes mixed-breed dogs (18.4%), Yorkshire Terriers (8.8%), Labrador Retrievers (7.9%), Golden Retrievers (5.0%), and Fox Terriers (3.4%). The most common tumors were interstitial cell tumors (n=227, 50.8%). In contrast, 107 (23.9%) seminomas, 80 (17.9%) Sertoli cell tumors, 19 (7.4%) mixed germ cell-sex cord-stromal tumors, and 26 (7.6%) testicular tumors developed from cryptorchid testes, which included 16 (61.5%) Sertoli cell tumors, 10 (38.5%) seminomas, and no interstitial cell tumors.

Conclusion: This study provides baseline information on the prevalence of canine testicular tumors in the described population, including the median age of each tumor type and overrepresented dog breeds. We further found that the most common scrotal testicular tumor was interstitial cell tumor, whereas Sertoli cell tumor was the most common in cryptorchid testicles.

Keywords: cryptorchid testes, interstitial cell tumors: mixed germ cell-sex cord-stromal tumors, seminomas, Sertoli cell tumors, testicular tumors.

Current status and advances of fish vaccines in Malaysia

Reveiw (Published online: 26-02-2022)

30. Current status and advances of fish vaccines in Malaysia

Mohd Syafiq Mohammad Ridzuan, Azila Abdullah, Rimatulhana Ramly, Nur Nazifah Mansor, Norazsida Ramli and Mohd. Firdaus-Nawi

Veterinary World, 15(2): 465-482

ABSTRACT

Fish diseases have a significant negative influence on the Malaysian aquaculture industry. Since the 1980s, the sector has grown in size, which has resulted in a rise in the prevalence of infectious outbreaks affecting both freshwater and marine cultured fish species. Demand for commercially available fish vaccinations is predicted to increase as infectious disease outbreaks continue to occur. In Malaysia, aquaculture vaccine research and development (R&D) are still in its infancy, with most efforts concentrating on producing vaccines against bacterial infections, most notably streptococcosis, vibriosis, and motile Aeromonas septicemia. Despite several attempts, no homegrown vaccine has been effectively introduced into the manufacturing pipeline to date. At the moment, only three imported aquatic vaccines have received full permission, a far cry from the 314 and 60 vaccines licensed in the poultry and porcine industries, respectively. This review will describe recent findings regarding the development of aquaculture vaccines for certain fish species and diseases in Malaysia. In our opinion, R&D on fish vaccines is critical to the aquaculture industry's viability.

Keywords: aquaculture, fish, fish diseases, Malaysia, vaccine.

Seroprevalence of sheeppox and goatpox virus in Asia and African continent: A systematic review and meta-analysis (Scientometrics)

Research (Published online: 25-02-2022)

29. Seroprevalence of sheeppox and goatpox virus in Asia and African continent: A systematic review and meta-analysis (Scientometrics)

Kuralayanapalya Puttahonnappa Suresh, Anenahalli Panduranga Bhavya, Chandan Shivamallu, Raghu Ram Achar, Ekaterina Silina, Victor Stupin, Shiva Prasad Kollur, Bibek Ranjan Shome and Sharanagouda S. Patil

Veterinary World, 15(2): 455-464

ABSTRACT

Background and Aim: Two endemic capripox infectious diseases, sheeppox (SP) and goatpox (GP) are common in Asia, Africa, and the Middle East. Sheep and goats, in general, are considered current assets of small and marginal farmers and have significant economic value in terms of meat, wool, and skin/hide production. Sheep and goat populations in India total 148.88 million and 74.26 million, respectively. Capripox caused US$ 2.3 million (Indian Rupee [INR] 105 million) in economic damages in Maharashtra (India) alone, and it took over 6 years for a flock to recover from the outbreak. The projected yearly loss at the national level is US$ 27.47 million (INR 1250 million). As a result, Capripox diseases put small and marginal farmers under much financial strain. The present study estimates the seroprevalence of SP and GP diseases in the Asian and African continents using systematic review and meta-analysis. The results of the study will help researchers and policymakers to understand the spatial and temporal distribution of the disease and its burden. In addition, the results are also helpful to design and implement location-specific prevention and eradication measures against these diseases.

Materials and Methods: Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines of Cochran collaborations were used for systematic review and subsequently meta-analysis were used. The literature was collected from various databases. Initial search string resulted in more than nine thousand articles for the period 2000 to 2020 using the different combinations of keywords and Boolean operators (or not) asterisk∗ and quotation marks. Out of 9398 papers, 80 studies were chosen for complete test reviews and quality bias evaluation using the inclusion and exclusion criteria. Finally, 21 articles were used for the meta-analysis. The statistical study employed fixed effects and random effects models using R.

Results: Seroprevalence of SP and GP was calculated using studies with a cumulative sample size of 4352, out of which sheep and goats' samples together contribute 48%, followed by sheep (32%) and goat (21%). The result of the meta-regression revealed that detection techniques had a significant impact on the overall effect size at 5% level (Qm=14.12). Subgroup analysis of polymerase chain reaction (PCR) test with samples was further grouped into two categories based on the median, and it revealed that 62% of samples used PCR as a detecting test followed by group-II.

Conclusion: From the study, it is concluded that SP and GP diseases are highly prevalent; hence, effective vaccines, proper education to farmers through extension activity, and transboundary disease movement restriction are necessary for the control and eradication of the disease.

Keywords: Asia, Africa, capripoxvirus, meta-analysis, seroprevalence, subgroup analysis.

Genomic structure of Bali cattle based on linkage disequilibrium and effective population size analyses using 50K single nucleotide polymorphisms data

Research (Published online: 25-02-2022)

28. Genomic structure of Bali cattle based on linkage disequilibrium and effective population size analyses using 50K single nucleotide polymorphisms data

Pita Sudrajad, Richi Yuliavian Kusminanto, Slamet Diah Volkandari and Muhammad Cahyadi

Veterinary World, 15(2): 449-454

ABSTRACT

Background and Aim: Bali Cattle (Bos j. javanicus) is a local breed originating in Indonesia, accounting for 32.3% of the total cattle population. To date, no studies of the genetic structure and demographic status of Bali cattle have been conducted, even though the breeding of Bali cattle has a long and unique history that is likely to have impacted its genetic diversity. Therefore, a study that used molecular breeding technologies to characterize the demography of Bali cattle would be timely. This study aimed to examine genome diversity in Bali cattle and estimate the linkage disequilibrium (LD) and effective population size (Ne) values in the cattle population.

Materials and Methods: In this study, we explored the population structure and genetic diversity of Bali cattle using genomic-level analyses. Our study primarily studied cattle that had been bred in livestock breeding centers since these breeds had subsequently spread throughout Indonesia. We focused on characterizing the genetic structure, determining the level of LD present, and estimating the Ne of the Bali cattle population. The genomic data used for this study were obtained from DNA samples of 48 Bali cattle collected at the Breeding Center of Bali Cattle as well as 54 genomic samples from Bali cattle collected elsewhere in Indonesia that had been used in recent publications. This genomic dataset included exclusively 50K single nucleotide polymorphisms (SNP) array (Illumina Bovine 50SNP bead chip, Illumina, USA) data.

Results: We found that the LD values of Bali cattle from the breeding center and those raised elsewhere were 0.48±0.43 and 0.39±0.40, respectively. Subsequently, the Ne value of Bali cattle from the breeding center and farmers was 151 and 96, respectively.

Conclusion: Our results suggest that the selection program of the breeding center is beneficial for maintaining the genetic diversity of Bali cattle.

Keywords: Bali cattle, effective population size, genomic data, genetic structure, linkage disequilibrium.

The first study on seroprevalence and risk factors of Neospora caninum infection in pregnant local cows from Northeast Algeria

Research (Published online: 25-02-2022)

27. The first study on seroprevalence and risk factors of Neospora caninum infection in pregnant local cows from Northeast Algeria

Besma Abdeltif, Safia Tennah, Salima Yamina Derdour, Asma Temim, Houda Boufendi and Farida Ghalmi

Veterinary World, 15(2): 442-448

ABSTRACT

Background and Aim: Neospora caninum is one of the most common infectious organisms worldwide that causes abortion in cattle. To the best of our knowledge, no previous studies have focused on N. caninum infection in the local Atlas brown cattle from Northeast Algeria. This study aimed to assess the importance of bovine neosporosis for causing abortion in Atlas brown cattle and to identify selected risk factors.

Materials and Methods: A case-control study was performed on 60 control farms and 30 case farms. We collected 650 blood samples from 650 pregnant cows from 90 farms in five Algerian provinces; Jijel, Skikda, Annaba, El-Tarf, and Souk- Ahras. Sera samples were analyzed for the presence of antibodies against N. caninum using enzyme-linked immunosorbent assays.

Results: The seroprevalence of N. caninum infection in the cows was 36.2% (95% confidence interval [CI]: 32.7-39.8) and in the farms was 81.1% (95% CI: 73.0-89.2). Risk factors found by multivariable logistic regression included: Presence of dogs (odds ratio [OR] 4.7, 95 CI 2.9-7.3); age ≥84 months (OR 4.9, 95 CI 2.8-8.3); Jijel region (OR 2.2, 95 CI 1.1-4.5); white (OR 2.5, 95 CI 1.4-4.4) and gray (OR 2.5, 95 CI 1.4-4.5) coat; moderate (OR 2.30, 95 CI 1.4-3.8) and bad (OR 3.1, 95 CI 1.8-5.3) hygiene; and second (OR 2.5, 95 CI 1.4-4.4); and last (OR 2.3, 95 CI 1.3-4.2) stage of pregnancy. Our case-control study showed no significant association between seropositivity of N. caninum and abortion at the farms level (OR 0.9, 95 CI 0.3-2.7). Similarly, there was no significant association between seropositivity of N. caninum and abortion at the individual level (OR 0.8, 95 CI 0.6-1.2).

Conclusion: This is the first study of N. caninum infection in pregnant local cows from Northeast Algeria. The prevalence rate of antibodies against N. caninum was high. Almost all risk factors studied for infection were significantly associated with seroprevalence. Our analysis showed no relation between N. caninum infection and abortion. Consequently, these local cows are resistant to abortion caused by N. caninum.

Keywords: Algeria, Neospora caninum, pregnant cattle, seroepidemiology.