scholarly journals Molecular Prevalence of Anaplasma marginale and Ehrlichia in Domestic Large Ruminants and Rhipicephalus (Boophilus) microplus Ticks From Southern Luzon, Philippines

2021 ◽  
Vol 8 ◽  
Author(s):  
Remil L. Galay ◽  
Carina R. Llaneta ◽  
Maria Karla Faye B. Monreal ◽  
Antero L. Armero ◽  
Arianne Bel D. Baluyut ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Water Buffalo ◽  
Anaplasma Marginale ◽  
Epidemiological Data ◽  
The Philippines ◽  
Boophilus Microplus ◽  
Economic Losses ◽  
Molecular Evidence ◽  
Blood Samples ◽  
Pcr Targeting

Anaplasmosis and ehrlichiosis are tick-borne rickettsial diseases that cause significant economic losses in the livestock industry worldwide. Although bovine anaplasmosis is known to be endemic in the Philippines, epidemiological data is fragmented. Moreover, little is known about bovine ehrlichiosis in the country. In this study, the prevalence of Anaplasma marginale and Ehrlichia in cattle and water buffalo from provinces in the southern part of Luzon, Philippines, was investigated through PCR. Blood samples from 620 animals comprised of 512 cattle and 108 water buffalo and 195 tick samples were subjected to nested PCR targeting the groESL gene of Anaplasmataceae. Positive samples were further subjected to another nested PCR and conventional PCR to amplify the A. marginale groEL gene and the Ehrlichia dsbA gene, respectively. Selected A. marginale-positive samples were also subjected to nested PCR targeting the msp5 gene. Regardless of the animal host, the overall prevalence in blood samples obtained was 51.9% for Anaplasmataceae, 43% for A. marginale, and 1.1% for Ehrlichia. No water buffalo were positive for Ehrlichia. Meanwhile, 15.9, 6.7, and 2% of the tick samples, all morphologically identified as Rhipicephalus (Boophilus) microplus, were positive for Anaplasmataceae, A. marginale, and Ehrlichia, respectively. Sequence analysis of selected A. marginale msp5 amplicons showed that the isolates from the region share 94–98% identity to reported A. marginale from other countries. The phylogenetic tree showed clustering of isolates in the region and a close relationship with A. marginale isolates from other countries. Sequences of Ehrlichia amplicons from cattle and ticks were 97–100% similar to reported Ehrlichia minasensis isolates. This study showed the high prevalence of A. marginale in Luzon, Philippines, and provided the first molecular evidence of E. minasensis in the country.

scholarly journals Molecular Epidemiology of Bovine Babesiosis in Punjab, Pakistan

2021 ◽  
Vol 49 ◽  
Author(s):  
Asif Masih ◽  
Azhar Rafique ◽  
Farhat Jabeen ◽  
Shabana Naz
Keyword(s):  
Risk Factors ◽  
Nested Pcr ◽  
Dairy Industry ◽  
Summer Season ◽  
Babesia Bovis ◽  
Economic Losses ◽  
Molecular Evidence ◽  
Conventional Pcr ◽  
Associated Risk Factors ◽  
Pcr Targeting

Background: Babesiosis is endemic in Pakistan and is one of the most important bovine diseases that causes huge economic losses and high mortality in young animals. This disease is transmitted by a protozoan parasite babesia which belongs to genus Babesia (Apicomplexa: Piroplasmida: Babesiidae). This disease is very much prevalent in summers followed by rainy season because humid environment is favorable for the growth of these parasites. An epidemiological and molecular study was conducted to unveil the prevalence and associated risk factors of Babesia bigemina (B. bigemina) and Babesia bovis (B. bovis) in selected districts i.e., Faisalabad, Toba Tek Singh and Jhang of Punjab, Pakistan.Materials, Methods & Results: A total of 518 (Cattle = 360, Buffalo = 158) blood samples were collected. The samples were analyzed by polymerase chain reaction (PCR) and nested PCR (n-PCR) targeting apocytochrome b-genes (CYTb). Chi-square test for univariate analysis was used to analyze the data. The overall prevalence in summer based upon microscopic analysis was 20.55% (37/180) and 13.92% (11/79) in cattle and buffaloes respectively and in winter was 8.80% (16/180), 5.06% (4/79)) in cattle and buffaloes respectively. The samples were further analyzed through conventional PCR (c-PCR) and nested PCR (nPCR). The overall results of conventional PCR in summer showed that 72 cows and buffaloes were infected with babesiosis. The conventional PCR based results of summer showed that prevalence of babesiosis was 29.44% (53/180) in cows and 24.05% (19/79) buffaloes. The results of cPCR during the winter season showed that 12.77% (23/180) and 13.92% (11/79) buffaloes were positive for babesiosis. The overall results of conventional PCR in winter showed that 34/259 cows and buffaloes were infected with babesiosis. On the other hand, the nested PCR results of summer season showed that the prevalence of babesiosis in cows was 32.22% (58/180) and 29.11% (23/79) in buffaloes. In total, 81 cows and buffaloes were infected with babesiosis during summer season. The nPCR results of winter showed that 15% (27/180) cows and 20.25% (16/79) buffaloes were infected with babesiosis. In total, 43 cows and buffaloes were infected with babesiosis. The results have shown that sensitivity of n-PCR is more as compared to conventional PCR. This study is the first molecular evidence of B. bigemina and B. bovis and its associated risk factors in Punjab province, Pakistan.Discussion: Dairy sector in Pakistan is one of the fastest growing sectors. Despite of remarkable growth, dairy industry is facing many problems one of them is tick borne diseases (TBDs). TBDs are more prevalent in tropical and subtropical areas of the world and leads to huge economic losses to dairy industry in terms of decreased milk, meat and wool production. Babesiosis is characterized by increased fever, decreased production, poor quality wool, anemia, hemoglobinuria, paleness of mucous membrane. The risk factors analysis of summer and winter data revealed that, adult animals were more prone to babesiosis (24.00%) [P = 0.032] and (8.50%) [P = 0.048]. In both seasons (summer and winter), females were more infected with babesiosis (20.19% and 8.17%) [P = 0.049 and P =0.021] as compared to males, high prevalence in females was might be due to that females were reared for longer period of time. Babesiosis was more occurred in non-cemented floor system (26.01% and 13.51%) [P = 0.028 and P = 0.044] in summer and winter, respectively. Disease was found more prevalent in closed housing system in summer and winter (27.27% and 10.93%) [P = 0.043 and P = 0.034] as compared to open housing. Weak animals were more infected with babesiosis (30.84%) [P = 0.045] and (12.80%) [P = 0.042] in summer and winter, as compared to healthy ones. The animals with high tick infestations were more suffered with babesia infection (25.49% and 13.34%) [P = 0.036 and P = 0.003] in both seasons as compared to less tick burden. Keywords: apocytochrome gene, babesiosis, bovine, nPCR, PCR, season.

scholarly journals Molecular Detection of Rickettsia spp. and Coxiella burnetii in Cattle, Water Buffalo, and Rhipicephalus (Boophilus) microplus Ticks in Luzon Island of the Philippines

2020 ◽  
Vol 5 (2) ◽  
pp. 54 ◽  
Author(s):  
Remil L. Galay ◽  
Melbourne R. Talactac ◽  
Bea V. Ambita-Salem ◽  
Dawn Maureen M. Chu ◽  
Lali Marie O. dela Costa ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Molecular Detection ◽  
Water Buffalo ◽  
The Philippines ◽  
Boophilus Microplus ◽  
Positive Sequence ◽  
Effective Control ◽  
Molecular Evidence ◽  
Cattle Ticks ◽  
Boophilus Microplus Ticks

Rickettsia and Coxiella burnetii are zoonotic, tick-borne pathogens that can cause febrile illnesses with or without other symptoms in humans, but may cause subclinical infections in animals. There are only a few reports on the occurrence of these pathogens in cattle and water buffalo in Southeast Asia, including the Philippines. In this study, molecular detection of Rickettsia and C. burnetii in the blood and in the Rhipicephalus (Boophilus) microplus ticks of cattle and water buffalo from five provinces in Luzon Island of the Philippines was done. A total of 620 blood samples of cattle and water buffalo and 206 tick samples were collected and subjected to DNA extraction. After successful amplification of control genes, nested PCR was performed to detect gltA of Rickettsia and com1 of C. burnetii. No samples were positive for Rickettsia, while 10 (cattle = 7, water buffaloes = 3), or 1.6% of blood, and five, or 1.8% of tick samples, were C. burnetii-positive. Sequence analysis of the positive amplicons showed 99–100% similarity to reported C. burnetii isolates. This molecular evidence on the occurrence of C. burnetii in Philippine ruminants and cattle ticks and its zoonotic nature should prompt further investigation and surveillance to facilitate its effective control.

scholarly journals Serological and molecular evidence of Brucella species in the rapidly growing pig sector in Kenya

2020 ◽  
Author(s):  
James Miser Akoko ◽  
Roger Pelle ◽  
Velma Kivali ◽  
Esther Schelling ◽  
Gabriel Shirima ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Epidemiological Data ◽  
Molecular Techniques ◽  
Molecular Evidence ◽  
Rt Pcr ◽  
Growing Pig ◽  
Pcr Targeting ◽  
The Rose

Abstract BackgroundBrucellosis is an emerging, yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi, Kenya were screened in parallel, using the Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp.. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 gene and real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes.ResultsA prevalence of 0.57% (n=4/700) was estimated using RBT. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR (n = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a multiple real-time PCR. ConclusionThe detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs indicate the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

scholarly journals First report and molecular characterization of Cryptosporidium spp. in humans and animals in Khartoum state, Sudan

2019 ◽  
Vol 12 (1) ◽  
pp. 183-189 ◽  
Author(s):  
Kaltoum Yagoub Adam ◽  
A. A. Ismail ◽  
M. A. Masri ◽  
A. A. Gameel
Keyword(s):  
Nested Pcr ◽  
Mammalian Species ◽  
Pcr Amplification ◽  
Health Concern ◽  
Economic Losses ◽  
Staining Procedure ◽  
Rrna Gene ◽  
Nested Polymerase Chain Reaction ◽  
Cryptosporidium Spp ◽  
Pcr Targeting

Background and Aim: Cryptosporidium is recognized to infect several mammalian species as well as humans, causing substantial economic losses and serious public health concern. Infected animals can be a source of environmental contamination and human infections. In general, the occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained. Materials and Methods: To provide molecular information about Cryptosporidium in animals and humans, both modified Ziehl-Neelsen (MZN) specific stain and molecular assay were used. Concentration techniques followed by three protocols of DNA extraction were carried out. After microscopic screening of 263 fecal samples (goats [n=197], cattle [n=12], sheep [n=12], and human [n=42]), 61 positive and 30 negative, randomly selected samples were used in nested polymerase chain reaction (PCR) targeting part of the 18S RNA. Results: Nested PCR amplification confirmed 91.8% (56/61) of microscopic-positive samples. 8.2% (5/61) of negative samples by PCR (positive by microscopy) were considered false negatives. Sequencing followed by alignment of the 14 isolates indicated that all samples were identical (100%) and belonged to Cryptosporidium parvum. Conclusion: MZN staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested PCR targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite.

scholarly journals Ringworm in calves: Risk factors, improved molecular diagnosis, and efficacy of Aloe vera gel extract for treatment

2019 ◽  
Author(s):  
Yasmine Hasanine Tartor ◽  
Wafaa M. I. El-Neshwy ◽  
Abdallah M. A. Merwad ◽  
Mohamed F. Abo El-Maati ◽  
Rehab E. Mohamed ◽  
...  
Keyword(s):  
Risk Factors ◽  
Nested Pcr ◽  
Aloe Vera ◽  
Effective Control ◽  
Economic Losses ◽  
Diagnostic Indices ◽  
Aloe Vera Gel ◽  
Hair Samples ◽  
Conventional Methods ◽  
Pcr Targeting

Abstract Background: Calves dermatophytosis is a major public and veterinary health problem worldwide due to its zoonotic potential and economic losses in cattle farms. However, it has lacked adequate attention; thereby for effective control measures it is worth determining ringworm prevalence, risk factors and direct sample nested-PCR diagnostic indices as compared to conventional methods for dermatophytes identification. Moreover, Aloe vera gel extract (AGE) phenolic composition and its in-vitro and in-vivo anti-dermatophytic activity in comparison to antifungal drugs were evaluated. Results: Of 760 examined calves, 55.79 % showed ringworm lesions. 84.91% were positive for fungal elements in direct microscopy, and 79.72% were culture positive. Trichophyton verrucosum was the most frequently identified dermatophytes (90.24%). Risk of dermatophytosis is high in 4-6 month than 1-month aged calves (60% versus 41%), in summer and winter compared to spring and autumn seasons (66% and 54% versus 48%). Poor hygienic conditions, intensive breeding system, animals raised for meat production, parasitic infestation, crossbreed, and newly purchased animals were statistically significant risk factors correlated with dermatophytosis. One-step PCR targeting conserved regions in the 18S and 28S genes achieved unequivocal identification of T. verrucosum and T. mentagrophytes in hair samples. Nested-PCR achieved an excellent performance in all tested diagnostic indices and increased the species-specific detection of dermatophytes by 20 % as compared to culture. Terbinafine and miconazole were the most active antifungal agents for dermatophytes. Gallic acid, caffeic acid, chlorogenic acid, cinnamic acid, aloe-Emodin, quercetin, and rutin are the major phenolic compounds of AGE identified by High-performance liquid chromatography (HPLC). These compounds increased and synergized the anti-dermatophytic activity of AGE. The treated groups showed significantly lower clinical scores than the control group ( P < 0.05). The calves were successfully treated with topical AGE (500 ppm) resulting in clinical and mycological cure within 14-28 day of the experiment. Conclusions: Implementation of nested-PCR assay providing a rapid diagnostic tool for dermatophytosis augments and complement the conventional methods for initiating targeted treatments of calves ringworm. The recognized anti-dermatophytic potential of AGE is advantageous countenance to commercial drugs to go used in therapeutics.

scholarly journals High genetic diversity of Anaplasma marginale infecting dairy cattle in northeastern Brazil

2021 ◽  
Vol 30 (4) ◽  
Author(s):  
José Gomes Pereira ◽  
Amanda Barbosa Garcia ◽  
Luiz Ricardo Gonçalves ◽  
Inalda Angélica de Souza Ramos ◽  
Maria do Socorro Costa Oliveira Braga ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Dairy Cattle ◽  
Anaplasma Marginale ◽  
The State ◽  
Amino Acid Sequences ◽  
Surface Proteins ◽  
Northeastern Brazil ◽  
High Genetic Diversity ◽  
Blood Samples ◽  
Pcr Targeting

Abstract Anaplasma marginale is an obligate intracellular Gram-negative bacterium found in ruminants’ erythrocytes and is the etiological agent of bovine anaplasmosis. The bacterium’s genetic diversity has been characterized based on sequences of major surface proteins (MSPs), such as MSP1α. The aim of the present study was to investigate the genetic diversity of A. marginale in cattle in the state of Maranhão, northeastern Brazil. To this end, 343 blood samples were harvested and subjected to iELISA assays using the recombinant surface protein MSP5. Out of 343 blood samples, 235 (68.5%) were randomly chosen and submitted to DNA extraction, qPCR and conventional PCR targeting the msp1α gene to determine amino acid sequences and classify the genotypes. The iELISA results showed 81.34% seropositivity (279/343), whereas qPCR revealed 224 positive samples (95.32%). Among these qPCR-positive samples, 67.4% (151/224) were also positive in the cPCR. Among the 50 obtained sequences, 21 strains had not been previously reported. Regarding the genotypes, H (26/50) and E (18/50) were identified most often, while genotypes F and C were only identified twice each and B and G once each. In conclusion, high prevalence and genetic diversity for A. marginale were observed in dairy cattle herds in the state of Maranhão.

scholarly journals Tick and Tick Borne Protozoan Diseases of Livestock in the Selected Hilly Areas of Bangladesh

2013 ◽  
Vol 1 (1-2) ◽  
pp. 60-63 ◽  
Author(s):  
UK Mohanta ◽  
Dr Anisuzzaman ◽  
MMH Mondal
Keyword(s):  
Rainy Season ◽  
Anaplasma Marginale ◽  
Boophilus Microplus ◽  
The Other ◽  
Seasonal Prevalence ◽  
Babesia Bigemina ◽  
Blood Samples ◽  
Other Hand ◽  
Amblyomma Testudinarium ◽  
One Year

To study the tick and tick borne protozoan diseases of livestock in the hilly areas of Bangladesh, an attempt was made to collect tick and blood samples from cattle, goat and gayal (Bos frontalis) from different areas of the three hill districts. In this study, two species of ticks namely, Boophilus microplus (92%) and Amblyomma testudinarium (21.6%) and two species of blood protozoa like Babesia bigemina (16.63%) and Anaplasma marginale (14.94%) were recorded. Seasonal prevalence of ticks was highest in summer (97%) in comparison to rainy (95%) and winter (86%) season. On the other hand, the seasonal prevalence of blood protozoa was highest in rainy season (45.45%) in comparison to summer (27.87%) and winter (16.55%). Again, animals aged more than 2 (two) years of age (52%) found to be more susceptible to blood protozoan diseases than animals aged between 1-2 years of age (33.97%). But none of the animals under one year of age were found to be infected with blood protozoan diseases. DOI: http://dx.doi.org/10.3329/ijarit.v1i1-2.13934 Int. J. Agril. Res. Innov. & Tech. 1 (1&2): 60-63, December, 2011

scholarly journals Serological and Molecular Evidence of Brucella Species in the Rapidly Growing Pig Sector in Kenya

2020 ◽  
Author(s):  
James Miser Akoko ◽  
Roger Pelle ◽  
Velma Kivali ◽  
Esther Schelling ◽  
Gabriel Shirima ◽  
...  
Keyword(s):  
Real Time ◽  
Epidemiological Data ◽  
Molecular Techniques ◽  
Molecular Evidence ◽  
Rt Pcr ◽  
Conventional Pcr ◽  
Brucella Infection ◽  
Growing Pig ◽  
Pcr Targeting

Abstract Background: Brucellosis is an emerging yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi region, Kenya were screened in parallel, using both Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 geneand real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes. Results: A prevalence of 0.57% (n = 4/700) was estimated using RBT; none of these samples was positive on cELISA. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR (n = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a real-time multiplex PCR. Conclusion: The detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs confirm the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

Molecular evidence of the reservoir competence of water buffalo (Bubalus bubalis) for Anaplasma marginale in Cuba

2018 ◽  
Vol 13 ◽  
pp. 180-187 ◽  
Author(s):  
Dasiel Obregón ◽  
Belkis G. Corona ◽  
José de la Fuente ◽  
Alejandro Cabezas-Cruz ◽  
Luiz Ricardo Gonçalves ◽  
...  
Keyword(s):  
Water Buffalo ◽  
Anaplasma Marginale ◽  
Bubalus Bubalis ◽  
Molecular Evidence

scholarly journals Serological and molecular evidence of Brucella species in the rapidly growing pig sector in Kenya

2020 ◽  
Author(s):  
James Miser Akoko ◽  
Roger Pelle ◽  
Velma Kivali ◽  
Esther Schelling ◽  
Gabriel Shirima ◽  
...  
Keyword(s):  
Real Time ◽  
Epidemiological Data ◽  
Molecular Techniques ◽  
Molecular Evidence ◽  
Brucella Species ◽  
Rt Pcr ◽  
Conventional Pcr ◽  
Growing Pig ◽  
Pcr Targeting

Abstract Background: Brucellosis is an emerging yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi region, Kenya were screened in parallel, using both Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 gene and real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes.Results: A prevalence of 0.57% (n = 4/700) was estimated using RBT; none of these samples was positive on cELISA. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR (n = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a real-time multiplex PCR.Conclusion: The detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs confirm the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

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