Economic and Social Impacts of COVID-19 on Animal Welfare and Dairy Husbandry in Central Punjab, Pakistan.

Studies on the impact of the COVID-19 pandemic on animal welfare and dairy husbandry in low-income countries are limited. We conducted a survey between February and June 2020 to evaluate the economic impact and animal health problems upon the pandemic. Participants were lead veterinarians from 14 dairy farms with herd size between 100 and 500 in Lahore. These farms were major suppliers of milk and dairy products to central Punjab, Pakistan. During the pandemic, 10 of the 14 dairy farms relied on feed mill concentrates to provide feeds to their herds. Half of the farms reported feed shortage due to lockdowns. Six (43%) dairy farms have witnessed a 7.5% shortage of dry feed intake. In seven (50%) farms, the body condition score decreased by 0.24 point. The body score reduction was significantly associated with depleted feed intake (P = 0.005). The veterinarians of 10 (71%) farms failed to gain access to essential veterinary medications, hampering the treatment of sick animals. Due to feed shortage and drug unavailability, daily milk production reduced by two litters per cow in the herd of five (35%) farms. The reduced feed intake was significantly associated with the decrease in milk production (P = 0.003), while numerous downstream milk-processing facilities were out of service during the pandemic, significantly reducing the profit of six (43%) dairy farms. Finally, our study showed that the dairy farming industry and animal welfare were critically affected by three aspects: feed shortage, inaccessibility to essential veterinary drugs, and a reduced consumer demand for dairy products.

Parasitic Mite Fauna in Asian Poultry Farming Systems.

The ubiquitous presence of hematophagous avian mites threatens the poultry industry in Asia and worldwide, adversely affecting the quality and quantity of eggs and poultry meat produced by affected flocks. This leads to considerable economic loss and welfare-related issues. The role of these blood-feeding arthropods as disease vectors is increasingly recognized as they may carry important zoonotic and epizootic pathogens. The poultry mites, Dermanyssus gallinae (Poultry Red Mite-PRM), Ornithonysus sylviarum (Northern Fowl Mite-NFM), and Ornithonyssus bursa (Tropical Fowl Mite-TFM) are endemic species across the Asian continent. In less frequency, scaly leg mite, depluming mite, and fowl cyst mite were documented. Considering global climate change and the popularity of backyard farming, the incidence of avian mite infestation is expected to increase as Asian production expands. The TFM may start to colonize sub-tropical nations where the seasonal temperature is comparable to tropical regions. Pyrethroids, organophosphates, carbamates, and macrocyclic lactones are licensed acaricides for use in China, Japan, and India. In recent years, the development of acaricide resistance has compromised the efficacy of chemical control measures. Several botanical acaricides based on plant and fungal constituents are being investigated. Judicious integration of multiple approaches to manage poultry mite infestation is advised. In this article, we review the prevalence, geographical distribution, zoonotic potential, and control measures of avian mites in poultry farms in Asia.

Circulation of Rickettsia species and rickettsial endosymbionts among small mammals and their ectoparasites in Eastern Slovakia.

Bacteria belonging to the genus Rickettsia are known as causative agents of vector-borne zoonotic diseases, such as spotted fevers, epidemic typhus and endemic typhus. Different species of ticks, mites and fleas could act as reservoirs and arthropod vectors of different pathogenic Rickettsia species. The aim of this work was to establish active surveillance of Rickettsia spp. in mites, ticks and fleas collected from small mammals (rodents and shrews) in Eastern Slovakia. A total of 964 animal ear biopsies, 871 mites, 667 ticks and 743 fleas were collected from small mammals in the Kosice region, Eastern Slovakia. All specimens were identified using specialized taxonomic keys, and were conserved in ethanol until DNA extraction was performed. After DNA extraction, identification of Rickettsia species was performed by PCR-based methods. The total prevalence of rickettsiae from ear biopsies was 4.6% (95% CI, 3.2-5.9), in tested mites 9.3% (95% CI, 7.4-11.2), 17.2% (95% CI, 14.3-20.1) in I. ricinus ticks and 3.5% (95% CI, 2.2-4.8) in fleas. Sequence analysis of the partial gltA gene and Rickettsia helvetica-, Rickettsia slovaca-, Rickettsia raoultii- species specific real-time PCR tests revealed the presence of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts. These pathogenic and symbiotic species were confirmed in the following ectoparasite species-Laelaps jettmari, Haemogamasus nidi, Laelaps agilis and Eulaelaps stabularis mites, Ixodes ricinus ticks, Ctenophthalmus solutus, C. assimilis and Megabothris turbidus fleas infesting host-Apodemus agrarius, A. flavicollis, Microtus arvalis and Myodes glareolus small mammals. These results confirm the circulation of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts in mites, ticks and fleas collected on small mammals in the Kosice region, Eastern Slovakia.

Association of mechanical cleaning and a liquid preparation of diatomaceous earth in the management of poultry red mite, Dermanyssus gallinae (Mesostigmata: Dermanyssidae).

The poultry red mite (PRM), Dermanyssus gallinae, is an ectoparasite of great importance related to poultry farms worldwide. Problems associated with its control have led to the search for alternative treatments, especially using inert dust, which has recently been introduced into liquid formulations. Therefore, the objective of this work was to evaluate the acaricidal activity of a liquid diatomaceous earth (DE) preparation in the laboratory and its association with mechanical cleaning (brushing) in the field. In the laboratory it was first determined that a concentration of 10% DE would be necessary for field applications. The field experiment was conducted in a commercial poultry house. The 10% DE liquid preparation was administered in three applications and associated to mechanical cleaning. Then, 42 days after the first DE application, a population reduction of 94.7% was observed in relation to the initial population of mites. These results confirm the efficiency of DE applications in association with mechanical cleaning already adopted in poultry houses as an alternative for the control of D. gallinae, which can also contribute to avoiding mite resistance to chemical acaricides.

The repellent efficacy of eleven essential oils against adult Dermacentor reticulatus ticks.

Dermacentor reticulatus ticks are among the most important arthropod vectors of zoonotic disease agents in Europe. Eleven essential oils, namely basil (Ocimum basilicum), bergamot (Citrus bergamia), clove bud (Syzygium aromaticum), citronella (Cymbopogon winterianus), creeping thyme (Thymus serpyllum), lavender (Lavandula angustifolia), lemon-scented gum (Corymbia citriodora), marjoram (Origanum majorana), peppermint (Mentha piperita), spearmint (M. spicata), and red thyme (Thymus vulgaris) were tested for repellency against adult D. reticulatus ticks at concentrations of 1 and 3%. Clove bud, creeping thyme and red thyme essential oils were the most efficient - repelling 83, 82 and 68% of ticks when diluted to 3%, respectively. The mixture of creeping thyme and citronella containing 1.5% of each showed higher repellency (91%) than individual essential oils at the concentration of 3%.

Isolation of the monooxygenase complex from Rhipicephalus (Boophilus) microplus - clues to understanding acaricide resistance.

The monooxygenase complex is composed of three key proteins, a cytochrome P450 (CYP), the cytochrome P450 oxidoreductase (CPR) and cytochrome b5 and plays a key role in the metabolism and detoxification of xenobiotic substances, including pesticides. In addition, overexpression of these components has been linked to pesticide resistance in several important vectors of disease. Despite this, the monooxygenase complex has not been isolated from the Southern cattle tick Rhipicephalus (Boophilus) microplus, a major disease vector in livestock. Using bioinformatics 115 transcriptomic sequences were analyzed to identify putative pesticide metabolizing CYPs. RACE-PCR was used to amplify the full length sequence of one CYP; CYP3006G8 which displays a high degree of homology to members of the CYP6 and 9 subfamilies, known to metabolize pyrethroids. mRNA expression levels of CYP3006G8 were investigated in 11 strains of R. microplus with differing resistance profiles by qPCR, the results of which indicated a correlation with pyrethroid metabolic resistance. In addition to this gene, the sequences for CPR and cytochrome b5 were also identified and subsequently isolated from R. microplus using PCR. CYP3006G8 is only the third CYP gene isolated from R. microplus and the first to putatively metabolize pesticides. The initial results of expression analysis suggest that CYP3006G8 metabolizes pyrethroids but further biochemical characterization is required to confirm this. Differences in the kinetic parameters of human and mosquito CPR in terms of NADPH binding have been demonstrated and could potentially be used to design species specific pesticides. Similar differences in the tick CPR would confirm that this is a characteristic of heamatophagous arthropods.

Prevalence of pathogenic bacteria in Ixodes ricinus ticks in Central Bohemia.

Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

Sympatric occurrence of Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks and Rickettsia and Babesia species in Slovakia.

Vojka nad Dunajom in the south-west of the Slovak Republic is a locality with sympatric occurrence of 3 species of ticks. This study investigated the spatial distribution of Dermacentor reticulatus, Ixodes ricinus, and Haemaphysalis concinna ticks in this area and determined the prevalence of Babesia and Rickettsia species in questing adults of these tick species considered as potential risk for humans and animals. Ticks were collected by blanket dragging over the vegetation from September 2011 to October 2012. All ticks were subjected to DNA extraction and individually assayed with PCR-based methods targeting the gltA, sca4, 23S rRNA genes of Rickettsia spp. and the 18S rRNA gene of Babesia spp. D. reticulatus was the dominant species occurring in this area (67.7%, n=600), followed by I. ricinus (31.8%, n=282) and H. concinna (0.5%, n=4) ticks. Rickettsial infection was determined in 10.8% (n=65) and 11.7% (n=33) of D. reticulatus and I. ricinus ticks, respectively. Babesia spp. infection was confirmed in 1.8% (n=11) of D. reticulatus and 0.4% (n=1) of I. ricinus ticks. DNA of 6 different pathogenic tick-borne species, Rickettsia helvetica, Rickettsia monacensis, Rickettsia slovaca, Rickettsia raoultii, Babesia canis, and Babesia venatorum were identified in this locality with sympatric occurrence of I. ricinus, D. reticulatus, and H. concinna ticks.

Present and future potential of plant-derived products to control arthropods of veterinary and medical significance.

The use of synthetic pesticides and repellents to target pests of veterinary and medical significance is becoming increasingly problematic. One alternative approach employs the bioactive attributes of plant-derived products (PDPs). These are particularly attractive on the grounds of low mammalian toxicity, short environmental persistence and complex chemistries that should limit development of pest resistance against them.Several pesticides and repellents based on PDPs are already available, and in some cases widely utilised, in modern pest management. Many more have a long history of traditional use in poorer areas of the globe where access to synthetic pesticides is often limited. Preliminary studies support that PDPs could be more widely used to target numerous medical and veterinary pests, with modes of action often specific to invertebrates.Though their current and future potential appears significant, development and deployment of PDPs to target veterinary and medical pests is not without issue. Variable efficacy is widely recognised as a restraint to PDPs for pest control. Identifying and developing natural bioactive PDP components in place of chemically less-stable raw or 'whole' products seems to be the most popular solution to this problem. A limited residual activity, often due to photosensitivity or high volatility, is a further drawback in some cases (though potentially advantageous in others). Nevertheless, encapsulation technologies and other slow-release mechanisms offer strong potential to improve residual activity where needed.The current review provides a summary of existing use and future potential of PDPs against ectoparasites of veterinary and medical significance. Four main types of PDP are considered (pyrethrum, neem, essential oils and plant extracts) for their pesticidal, growth regulating and repellent or deterrent properties. An overview of existing use and research for each is provided, with direction to more extensive reviews given in many sections. Sections to highlight potential issues, modes of action and emerging and future potential are also included.

Immune responses of the domestic fowl to Dermanyssus gallinae under laboratory conditions.

There appear to be few reports in the literature regarding the host-poultry red mite (Dermanyssus gallinae) immunological relationship, despite the negative impact D. gallinae can have on both bird welfare and egg production, as well as its potential to act as a reservoir of zoonotic infections. The current study investigated the effect of either continuous infestation (CI) for 22 days or repeated infestation (RI) for four 24-h periods 7 days apart with D. gallinae on humoral immunity (IgM and IgY) and Th1/Th2 cytokine mRNA expression in peripheral blood mononuclear cells (PBMC) compared to non-infested controls. Serum IgY levels and IgM concentration were significantly higher in CI than RI and control birds although Th1 and Th2 mRNA expression in PBMC did not differ significantly between groups. D. gallinae appeared to modify reproductive behaviour and progeny survival following successive feeding events. In the RI group, the proportion of eggs/mite was significantly higher (P < 0.05) after first infestation than later infestations while larval/nymphal mortality was significantly higher (P < 0.05) after the first two infestations than later infestations. These data suggest that D. gallinae might adopt a feeding strategy of minimal host interference while D. gallinae could determine host immune status via nymphal/larval survival rates. Further research is required to better understand the host immunomodulation or avoidance strategy of D. gallinae as well as whether the mite is able to determine host immunocompetence perhaps using progeny survival.

The poultry red mite (Dermanyssus gallinae): a potential vector of pathogenic agents.

The poultry red mite, D. gallinae has been involved in the transmission of many pathogenic agents, responsible for serious diseases both in animals and humans. Nowadays, few effective methods are available to control the ectoparasite in poultry farms. Consequently, this is an emerging problem which must be taken into account to maintain good health in commercial egg production. This paper addresses the vector capacity of the ectoparasite with special emphasis on salmonellae, pathogenic agents responsible for many of the most important outbreaks of food-borne diseases worlwide. It has been experimentally shown that D. gallinae could act as a biological vector of S. enteritidis and natural carriage of these bacteria by the mite on poultry premises has also been reported. It was also found that D. gallinae carried other pathogens such as E. coli, Shigella sp., and Staphylococcus, thus increasing the list of pathogenic agents potentially transmitted by the mite.

Endosymbiotic bacteria living inside the poultry red mite (Dermanyssus gallinae).

This study investigated the endosymbiotic bacteria living inside the poultry red mite collected from five samples of one commercial farm from the UK and 16 farms from France using genus-specific PCR, PCR-TTGE and DNA sequencing. Endosymbiotic bacteria are intracellular obligate organisms that can cause several phenotypic and reproductive anomalies to their host and they are found widespread living inside arthropods. The farm sampled from the UK was positive for bacteria of the genera Cardinium sp. and Spiroplasma sp. From France, 7 farms were positive for Cardinium sp., 1 farm was positive for Spiroplasma sp., 1 farm was positive for Rickettsiella sp. and 2 farms were positive for Schineria sp. However, it was not possible to detect the presence of the genus Wolbachia sp. which has been observed in other ectoparasites. This study is the first report of the presence of endosymbionts living inside the poultry red mite. The results obtained suggest that it may be possible that these bacterial endosymbionts cause biological modifications to the poultry red mite.

Variation in chemical composition and acaricidal activity against Dermanyssus gallinae of four eucalyptus essential oils.

The results of this study suggest that certain eucalyptus essential oils may be of use as an alternative to synthetic acaricides in the management of the poultry red mite, Dermanyssus gallinae. At a level of 0.21 mg/cm(2), the essential oil from Eucalyptus citriodora achieved 85% mortality in D. gallinae over a 24 h exposure period in contact toxicity tests. A further two essential oils from different eucalyptus species, namely E. globulus and E. radiata, provided significantly (P < 0.05) lower mite mortality (11 and 19%, respectively). Notable differences were found between the eucalyptus essential oils regarding their chemical compositions. There appeared to be a trend whereby the essential oils that were composed of the fewer chemical components were the least lethal to D. gallinae. It may therefore be the case that the complexity of an essential oil's chemical make up plays an important role in dictating the toxicity of that oil to pests such as D. gallinae.

The application of PCR and reverse line blot hybridization to detect arthropod-borne hemopathogens of dogs and cats in Trinidad.

Arthropod-borne diseases are important causes of morbidity and mortality of companion animals in Trinidad. As clinical signs are vague, more sensitive methods to diagnose these diseases based on the polymerase chain reaction (PCR) followed by reverse line blot hybridization (RLB) of amplified products are being developed. An RLB of 14 oligonucleotide probes coupled with polymerase chain-amplified regions of 16S rRNA or 18S rRNA genes of hemoparasites from cats and dogs detected Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, feline mycoplasmas ("Candidatus Mycoplasma haemominutum,"Mycoplasma haemofelis), and some unknown species within the Babesia/Theileria group and the Anaplasma/Ehrlichia tribe. Amplified products were obtained from blood samples collected from 348 dogs and 15 cats. Overall, hemopathogen DNA was detected in 92 (26.4 per cent) dogs and six (40.0 per cent) cats. E. canis (49, 14.1 per cent) and feline mycoplasma (5, 33.3 per cent) DNA were most frequently identified in dogs and cats, respectively. B. canis vogeli (1, 6.7 per cent) and E. canis (1, 6.7 per cent) were also detected in cats. Mixed infections of Anaplasma/Ehrlichia sp. and Babesia sp. were present in five (1.4 per cent) dogs, while mixed infections of the feline mycoplasmas were present in two (13.3 per cent) cats, one of which was also positive for E. canis. Pyrexia was significantly associated with a positive RLB result in dogs (P= 0.00, chi(2), 1 df). This is the first reported application of macro-arraying techniques to detect arthropod-borne hemopathogens of companion animals in the Americas and the first detection of DNA of B. canis vogeli and E. canis in cats in Trinidad.

Use of plant-derived products to control arthropods of veterinary importance: a review.

The use of synthetic products in veterinary pest management is becoming increasingly problematic. Issues, including pest resistance, product withdrawal, undesirable environmental persistence, and high mammalian toxicity associated with synthetic pesticides, are driving research to identify new pest management approaches. One approach employs the repellent/toxic effects of plant-derived products (PDPs). Several pesticides based on PDPs are already available in some areas of pest management. This review highlights instances in which such products have been used with success against pests of domestic animals, livestock, apiculture, and poultry.

Immunological control of the poultry red mite.

In the current study whole poultry red mite antigens were extracted and birds were immunized subcutaneously with either antigen in adjuvant (antigen group) or PBS in adjuvant (control group). Immune responses of birds following immunization were investigated by ELISA and Western blotting, while vaccine efficacy was assessed by feeding of red mites on birds. Immunized birds showed a significant (P < 0.05) increase in IgY titers after immunization compared to controls, while immunoglobulin A (IgA) and IgM did not change significantly. However, the antigen group had a generally higher increase in all immunoglobulin titers compared to the controls. Western blotting identified a number of protein bands at different molecular weights, although these were not different between treatments. PCR analysis of whole mite protein identified bacterial DNA that might have confounded immunological data. In addition, there was a trend toward reduced survival rate of red mites feeding on antigen-immunized birds, but the difference was not statistically significant compared to controls. This study demonstrates the potential for somatic red mite antigens to stimulate an antibody-mediated immune response, although this response did not confer protection to birds.

The application of PCR and reverse line blot hybridization to detect arthropod-borne hemopathogens of dogs and cats in Trinidad.

Arthropod-borne diseases are important causes of morbidity and mortality of companion animals in Trinidad. As clinical signs are vague, more sensitive methods to diagnose these diseases based on the polymerase chain reaction (PCR) followed by reverse line blot hybridization (RLB) of amplified products are being developed. An RLB of 14 oligonucleotide probes coupled with polymerase chain-amplified regions of 16S rRNA or 18S rRNA genes of hemoparasites from cats and dogs detected Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, feline mycoplasmas ("Candidatus Mycoplasma haemominutum,"Mycoplasma haemofelis), and some unknown species within the Babesia/Theileria group and the Anaplasma/Ehrlichia tribe. Amplified products were obtained from blood samples collected from 348 dogs and 15 cats. Overall, hemopathogen DNA was detected in 92 (26.4%) dogs and six (40.0%) cats. E. canis (49, 14.1%) and feline mycoplasma (5, 33.3%) DNA were most frequently identified in dogs and cats, respectively. B. canis vogeli (1, 6.7%) and E. canis (1, 6.7%) were also detected in cats. Mixed infections of Anaplasma/Ehrlichia sp. and Babesia sp. were present in five (1.4%) dogs, while mixed infections of the feline mycoplasmas were present in two (13.3%) cats, one of which was also positive for E. canis. Pyrexia was significantly associated with a positive RLB result in dogs (P= 0.00, chi(2), 1 df). This is the first reported application of macro-arraying techniques to detect arthropod-borne hemopathogens of companion animals in the Americas and the first detection of DNA of B. canis vogeli and E. canis in cats in Trinidad.

Study on Theileria lestoquardi antigens as potential vaccine candidates.

Theileria lestoquardi is the causative agent of malignant theileriosis of sheep and goats, causing morbidity and mortality in these animals worldwide. Western blot analysis based on T. lestoquardi schizont antigens was carried out using sera collected from Iranian sheep, which had been immunized with T. lestoquardi schizont-infected cells. The results of Western blot analysis demonstrated that schizont-immunized animals produced antibodies reacting with protein bands at 73, 42, 20, 14, and 12 kDa. Comparison of the results of the current Western blotting test with earlier studies of Theileria spp. revealed two immunogenic schizont proteins with molecular weights of 73 and 42 kDa shared between T. annulata and T. lestoquardi. Two other proteins with molecular weights of 14 and 12 kDa have not been previously found in other Theileria species. Our results suggest that the 73-kDa protein could be a potential vaccine candidate and that the 14- and 12-kDa proteins could be considered as diagnostic antigens.

The poultry red mite Dermanyssus gallinae as a potential carrier of vector-borne diseases.

The poultry red mite Dermanyssus gallinae is an obligatory blood-sucking parasite that is considered to be one of the most important ectoparasites in the poultry industry, mainly because it is responsible for important economic losses, leads to a reduction of welfare of laying hens, and may pose a disease risk to humans. As a result of these problems, much of the current research on this parasite targets new methods of control. Less attention has been paid to the importance of D. gallinae as a carrier of vector-borne diseases. Some authors have mentioned the possible involvement of D. gallinae in the transmission (both in vitro and directly isolated from the mites) of viral and bacterial agents. Our research group has demonstrated the presence of Mycobacterium spp. within D. gallinae. DNA coding for Mycobacterium spp. was successfully amplified from unfed adult D. gallinae, larvae, and eggs by using reverse transcription-polymerase chain reaction targeting the 16S rRNA gene. The results have suggested the possible transovarial and transstadial transmission of pathogens by D. gallinae.