Prevalence of Multidrug-Resistant Pseudomonas aeruginosa Isolated from Dairy Cattle, Milk, Environment, and Workers' Hands.

Pseudomonas aeruginosa is an opportunistic pathogen causing severe infection in animals and humans. This study aimed to determine the ecological distribution and prevalence of multidrug-resistant (MDR) P. aeruginosa isolated from dairy cattle, the environment, and workers' hand swabs. Samples (n = 440) were collected from farms and households (n = 3, each). Rectal swabs, udder skin swabs, milk, workers' hand swabs, feed, water, water sources, and beddings were collected. Samples were subjected to the bacterial identification of P. aeruginosa via 16S rRNA. Antimicrobial resistance (AMR) was detected either phenotypically using an antibiotic susceptibility test or genotypically with AMR resistance genes (ARGs) such as drfA, sul1, and ermB. P. aeruginosa was detected on dairy farms and households (10.3-57.5%, respectively), with an average of 23.2%. The resistance of dairy farm strains was observed against sulfamethoxazole, imipenem, cefepime, piperacillin-tazobactam, and gentamycin (100%, 72.7%, 72.7%, 68.8%, and 63.3%, respectively). Meanwhile, the resistance of household strains was observed against sulfamethoxazole, imipenem, amoxicillin, gentamicin, cefepime, and erythromycin by 91.3%, 82.6%, 75.4%, 75.4%, 68.1%, and 63.8%, respectively. The susceptibility of farm strains was detected against norfloxacin, ciprofloxacin, and levofloxacin (90.9%, 84.8%, and 72.7%, respectively). Meanwhile, the susceptibility of household strains was detected against ciprofloxacin, amikacin, and norfloxacin (100%, 84.1%, and 72.5%, respectively). About 81.4% of P. aeruginosa strains were MDR. ARGs (drfA, sul1, and ermB) were detected in farm strains (48.5%, 72.7%, and 24.4%, respectively) and household strains (50.7%, 72.5%, and 47.8%, respectively). Almost all P. aeruginosa had MAR over 0.2, indicating repeated application of antibiotics. P. aeruginosa prevalence was fivefold higher in households than on farms. MDR strains were higher amongst household strains than farm strains.

Serological Investigation and Epidemiological Analysis of Bovine Leptospirosis in Egypt.

Bovine leptospirosis is a bacterial zoonotic disease of worldwide distribution. Little information is available regarding the occurrence of the disease in the Nile Delta provinces, Egypt. The present study investigated the seroprevalence of leptospirosis among cattle from Dakahlia province, Northern Egypt, and identified the individual variables factors associated with infection. To this end, a total of 600 serum samples from cattle of small stakeholders with various clinical manifestations possibly associated with leptospirosis were collected from different localities across Dakahlia province, Egypt. Sera were examined serologically via ELISA to investigate the occurrence of the disease among animals. Chi-square test and multivariable logistic regression analyses were applied to determine the association between hypothesized risk factors and the disease. Interestingly, our findings showed that 39.33% of the examined sera were positive for Leptospira antibodies, with significant differences among different localities. In addition, statistical analysis showed significant differences among age groups. Notably, the highest prevalence rate (22%) was observed in those aged between 3 and 5 years (p < 0.0001), whereas the lowest prevalence (2.66%) was reported in cattle <1 year old (p < 0.0001). Moreover, females had a significantly higher prevalence rate (35.33%) than males (4%) (p < 0.0001). Furthermore, our results showed significant differences in the occurrence of infection and reported clinical signs (p < 0.0001). Multivariable logistic regression identified repeated breeder and drop milk yield as the best predictors for prediction of ELISA results and linear discriminant analysis (LDA) model showed that overall classification accuracy of ELISA result using clinical signs and demographic data as predictors was 70.7%. The current study concluded a relative high prevalence of leptospirosis among cows bred in movable herds and households in the studied area and that age, repeated breeder and drop milk yield can be considered major risk factors associated with infection.

Acaricidal Properties of Four Neem Seed Extracts (Azadirachta indica) on the Camel Tick Hyalomma dromedarii (Acari: Ixodidae).

Tick infestation remains one of the major health problems that affect the productivity and comfort of camels. The control of ticks mainly relies on using chemical acaracides. Limited information is available on the potential benefits and activity of various neem extracts on Hyalomma ticks. The present study investigated the acaricidal activity of neem seed extracts at different concentrations against developmental stages of the camel tick Hyalomma dromedarii in comparison to Butox and diazinon. The acaricidal activity of three extracts, namely, hexane extract (HE), methyl chloride extract (MCE), and methanol extract (ME), of neem seeds (Azadirachta indica) were tested at varying concentrations of 5, 10, 15, and 20% on engorged H. dromedarii female ticks at days 1, 3, 5, 7, 12, 16, 20, 28, 37, and 43 after treatment (DPT). Interestingly, results of applying different neem seed extracts to engorged H. dromedarii female ticks showed that the most effective extract was hexane at concentration 20%, causing 100% mortality at 1st day post-application, while methanol extract at 20% and dichloromethane extract at 20% caused the death of all ticks at 28th day posttreatment as compared to Butox® 5.0 and Diazinon-60, which resulted in mortality of all ticks at 3 and 5 DPT, respectively. In addition, no mortality was reported with the application of aqueous extract (AE), which served as the control group. Furthermore, the neem hexane extract exhibited high efficacy against reproductive performance of female ticks, whereas no fertility or oviposition was reported at all of their concentrations. Additionally, no hatchability occurred using all neem extracts, except the aqueous extract, which showing no effect. In the present study, larvae responded more rapidly to the plant extracts, whereas mortality of all larvae was recorded at 24 h after treatment with 5% hexane. Taken together, this study pointed out that the acaricidal effect of hexane extract of neem seeds was more effective and could be economically used for controlling H. dromedarii ticks.

Parasitological, Molecular, and Epidemiological Investigation of Cryptosporidium Infection Among Cattle and Buffalo Calves From Assiut Governorate, Upper Egypt: Current Status and Zoonotic Implications.

Details about the epidemiological patterns and real contributions of different reservoir animals in maintaining the transmission cycle of Cryptosporidium spp. in Upper Egypt remain lacking. This study was designed to investigate the occurrence of Cryptosporidium spp. in cattle and buffalo (n = 608) from Upper Egypt. The parasite for the resulting positive samples by fecal examination was molecularly identified using nested PCR targeting the small subunit rRNA. Moreover, several explanatory variables, including animals' age, sex, condition, seasonal variations, were examined to describe the epidemiological pattern of the disease. Interestingly, the fecal examination revealed that 33.55% (204/608) of the animals under study were infected with Cryptosporidium, including 38.27% among cattle and 28.16% among buffalo. The parasite was molecularly identified using nested PCR, and their amplicons were identified in almost all fecal samples using microscopy (202/204). According to age as an individual variable factor, the infection rates of Cryptosporidium spp. in cattle calves with ages of <1, 1-3, and >3 months were 39.13, 34.04, and 54.54%, respectively. Meanwhile, in buffalo calves, the occurrence rates were 28.57, 27.27, and 29.41%, respectively. Regarding sex, female cattle calves were more susceptible to Cryptosporidium infection (51.28%) than males (26.19%) (p < 0.05), whereas male buffalo calves had a higher infection rate (32.25%) than females (25%). According to seasonal variations, the infection rates of Cryptosporidium spp. in cattle calves during spring, summer, autumn, and winter were 42.11, 30.43, 30, and 52.63%, respectively. In contrast, lower infection rates of 30, 21.42, 23.52, and 35% were reported in buffalo calves during spring, summer, autumn, and winter, respectively. The rate of infection was 45.16% in diarrheic cattle calves and 15.78% in non-diarrheic ones (p < 0.05). Meanwhile, the infection rate was 33.96% in diarrheic buffalo calves and 11.11% in non-diarrheic ones (p < 0.05). This study reported a higher occurrence of Cryptosporidium infection among the animals under study and revealed that buffalos and cattle can contribute to maintaining the transmission cycle of this zoonotic parasite in Upper Egypt.

Epidemiological, Morphological, and Morphometric Study on Haemonchus spp. Recovered From Goats in Egypt.

Goats can be infected by multiple groups of external and internal parasites. Haemonchus spp. are among abomasal parasites that can result in higher mortality and several considerable economic losses in goats. Early detection of parasites and better understanding of the major risk factors associated with infection are among the main strategies for controlling the infection. Considering this, information on hemonchosis in goats from Egypt, and the contribution of goats in the maintenance of the epidemiological foci of the disease is limited. This study investigated the prevalence of Haemonchus species among 240 abomasum samples collected during postmortem examination of goat carcasses from Assiut Governorate, Egypt. Moreover, the association of the major risk factors to describe the epidemiological pattern of the disease was explored. This study demonstrated that 16.66% of abomasa samples harbored Haemonchus species. Additionally, age, sex, and sampling season were the most significant risk factors associated with infection. Following the variable factors under study, goats aged 1 year or older were at higher risk, with an infection rate of 22.14% (31 of 140), than those younger than 1 year (9%) [p = 0.008; odds ratio (OR) = 2.87; 95% confidence interval (CI), 1.30-6.35]. The infection rate was 25% (19 of 76) in males and 12.8% (21 of 164) in females [p = 0.024; odds ratio (OR) = 2.26; 95% confidence interval (CI), 1.13-4.53]. Moreover, the exposure to infection was higher in summer (22.22%) than in winter (8.33%) (p = 0.007; odds ratio (OR) = 0.318; 95% confidence interval (CI), 0.139-0.725). More importantly, three species of the parasite-Haemonchus contortus, Haemonchus placei, and Haemonchus longistipes-were identified for the first time, and the confirmation of the identification and morphological characterization of the worms was performed using light microscopy and SEM. Collectively, this study reveals interesting epidemiological, morphological, and morphometric findings associated with the occurrence of hemonchosis among goats in Egypt. This study suggests further research for exploring the major circulating species of the parasite in Egypt, which is mandatory for controlling the disease.

SPECIFIC MOLECULAR DETECTION OF PIROPLASMS AND CHARACTERIZATION OF ß-TUBULIN FOR A NOVEL BABESIA SPECIES IN SIKA DEER (CERVUS NIPPON YESOENSIS).

Piroplasms, which include Babesia spp. and Theileria spp., are protozoan parasites carried by ticks and commonly cause disease in animals and humans. Those caused by Babesia spp. manifest as fever, anemia, and hemoglobinuria, while Theileria spp. can lead to high fever, diarrhea, and lymphadenopathy. Recently, Theileria capreoli and an undescribed Babesia sp. were detected for the first time in sika deer (Cervus nippon yesoensis) from Hokkaido; however, there is limited information available on their epidemiology in Japan. Here, a touchdown polymerase chain reaction and reverse line blot hybridization were used to perform an epidemiological survey of T. capreoli and Babesia sp. using blood samples from 82 sika deer in Hokkaido, Japan. This was followed by partial sequencing and phylogenetic analysis of the 18S rRNA and ß-tubulin genes to characterize both piroplasm species. A total of 43 (52.4%) and 3 (3.7%) of the sika deer were positive for T. capreoli and Babesia sp., respectively. The ß-tubulin gene partial sequences for Babesia sp. were distinct from those of Babesia spp. in GenBank. Phylogenetic analysis showed that the unknown Babesia sp. is more closely related to B. bigemina and B. ovata than other Babesia spp. based on the ß-tubulin gene. Further studies are required to understand the ecology of these tick-borne pathogens in Japan.

Molecular investigation on the presence of canine parvovirus in Egypt.

Canine parvovirus type 2 (CPV-2) causes a highly contagious gastroenteritis disease of dogs and wild canids. To investigate the CPV-2 prevalence in Dakahlia Governorate, Egypt, a total of 50 fecal swabs were collected from suspected diseased dogs during 2016-2017. Out of 50 collected samples, 35 samples (70 %) presented positive results for CPV-2 using immuno-chromatography (IC) as a rapid test. CPV-2DNA was detected in 42 samples (84 %) by using polymerase chain reaction (PCR). The frequencies of CPV-2 were significantly higher in German shepherd breed (46 %; 23/50) and in age groups less than 6 months (76%; 38/50). We evaluated the breed, age, sex, rapid test results and clinical signs as predictors for classification of animal status into infected and not infected. The best predictors for classification process were rapid test result and clinical signs. Both CPV-2b and CPV-2c subtypes were detected by CPV2-VP2 gene sequences analysis. Deduced amino acid sequences alignment showed substitutions at 3 sites (Arg453Pro, Ala574Glu and Gln457Leu). Further investigations are needed to reveal the genetic and antigenic relation between field and vaccinal strains of CPV-2 in Egypt.

Utilizing attached hard ticks as pointers to the risk of infection by Babesia and Theileria species in sika deer (Cervus nippon yesoensis), in Japan.

Ticks are hematophagous ectoparasites that have a significant impact on their animal hosts. Along with mosquitoes, they are the main arthropod vectors of disease agents in domestic animals, wildlife and humans. To investigate the occurrence and prevalence of piroplasmids in ticks, DNA was extracted from 519 hard ticks collected from 116 hunted Hokkaido sika deer (Cervus nippon yesoensis). The success of the DNA extraction was confirmed by touchdown PCR targeting the mitochondrial 16S rDNA gene of ticks. Touchdown PCR and reverse line blot (RLB) hybridization targeting the 18S rRNA gene were used to detect 14 piroplasm species. All hard ticks parasitizing Hokkaido sika deer were identified as belonging to the genera Ixodes and Haemaphysalis. In total 163 samples (31.4%) were positive for Babesia and Theileria spp. among tick species according to RLB hybridization. Tick DNA hybridized to the oligonucleotide probes of Theileria sp. Thrivae (27.0% of ticks; 140/519), Theileria capreoli (10.6%; 55/519), Babesia divergens-like (1.7%; 9/519), Babesia sp. (Bab-SD) (0.6%; 3/519), Babesia microti U.S. (0.4%; 2/519), and B. microti Hobetsu (0.4%; 2/519). The partial sequencing and phylogenetic analyses of the 18S rRNA gene confirmed the RLB hybridization results. Further investigations are needed to reveal the epidemiology and respective vectors of these pathogens.

Epidemiological and Histopathological Investigation of Sarcoptic Mange in Camels in Egypt.

Mange has been considered one of the most common parasitic infestations among camels. It adversely impacts animal productivity and poses a risk to human health. Given the scarcity of available data about mange in camels, the current study focused on the prevalence of camel mange and its associated risk factors in Aswan Governorate, Egypt. Towards this end, a general visual inspection was conducted on camels (N = 210) in different markets and slaughterhouses in Aswan Governorate. Skin scrapings from suspect infected camels were also examined microscopically. Importantly, these findings were further checked and confirmed by histopathology on samples from suspected cases collected post-slaughter in abattoirs. The possible risk-associated factors, which include the camel's age, sex and sampling season, were recorded and statistically analyzed. Interestingly, the data showed that a total of 100 camels (47.6%) were found exclusively infested by sarcoptic mange. Furthermore, the predominant histopathological changes included burrowing tunnel of mites in the skin, hyperkeratosis and acanthosisconsis of the epidermis, while the dermis showed hemorrhage, mononuclear inflammatory cell infiltration around the blood vessels and perifolliculitis. These major histopathological findings are consistent with sarcoptic mange. Furthermore, the statistical analysis of the possible associated risk factors, camel's age (p = 0.006), gender (p = 0.032) and sampling season (p = 0.004), were all found to be significantly affected and related to the disease. In this regard, camels ≥2 years old were found at higher risk of infection (odds ratio (OR) = 2.75; 95% confidence interval (CI), 1.345 to 5.604) versus younger animals (OR = 0.36; 95 CI, 0.1784 to 0.743). Females had higher odds of exposure (OR = 2.02; 95% CI, 1.096 to 3.708) compared to males (OR = 0.50; 95% CI, 0.269 to 0.912). Moreover, the exposure to infection was reported higher in winter (OR = 2.30; 95% CI, 1.297 to 4.098) than in summer (OR = 0.43; 95% CI, 0.244 to 0.771). Collectively, our data provide novel epidemiological and histopathological support for sarcoptic mange being widespread among camels in the studied area. Sarcoptic mange is extremely contagious and zoonotic. Therefore, our baseline investigation indicates an urgent need for additional multicenter-studies to investigate the occurrence of this disease in camels and humans combined with the appropriate control measures of camel importation for combating this disease.

Molecular identification and characterization of piroplasm species in Hokkaido sika deer (Cervus nippon yesoensis), Japan.

Babesia and Theileria species are tick-borne protozoan parasites that have a veterinary and zoonotic importance. In order to investigate the prevalence and genetic diversity of these parasites, a total of 269 sika deer blood DNA samples collected from Hokkaido, Japan, were examined for Babesia and Theileria species by touch-down PCR targeting the 18S rRNA gene. Reverse line blot (RLB) hybridization was then used to detect 12 piroplasm species. The results revealed that 95.5% (257/269), 94.1% (253/269), 14.1% (38/269), 87.7% (236/269) and 11.5% (31/269) of the examined PCR products hybridized with the probes which were designed to detect all Babesia and Theileria spp., all Theileria spp., all Babesia spp., Theileria sp. Thrivae and Babesia divergens-like, respectively. The 18S rRNA gene partial sequences were divided into Theileria sp. Thrivae, T. capreoli, B. divergens-like and an undescribed Babesia species. This study showed the first detection of the undescribed Babesia sp. from Japan. Therefore, more studies are required to understand the ecology of the newly detected tick-borne pathogens in Hokkaido.