Sensitive, specific, and rapid on-site detection of calf diarrhea pathogens using the RPA-CRISPR/Cas 12a assay
Yan Wang, Yun Diao, Yun Diao
et al.
Calf diarrhea is a common gastrointestinal disease that usually occurs within one month of birth. The disease causes the greatest economic losses to the cattle industry. Currently, a variety of diagnostic methods have been developed for calf diarrhea infections. However, existing methods are still unsatisfactory in terms of sensitivity, specificity, simplicity, cost, and speed.To provide a more sensitive, specific, simpler, and faster detection method, we recently developed an RPA-CRISPR/Cas12a assay that can detect BVDV, BCoV, BRV, and ETEC infections in cattle on-site. Testing for each pathogen is performed in a single test tube, without the need to open the tube in the middle, and can be completed in under 50 minutes.The RPA-CRISPR/Cas12a assay can detect BVDV, BCoV, BRV, and ETEC at concentrations of at least 10 copies/μL. The RPA-CRISPR/Cas12a assay does not produce false-positive results due to the presence of other pathogens. The sensitivity of BCoV, BRV, and ETEC in the RPA-CRISPR/Cas12a quadruple assay is equivalent to that of single qPCR. The sensitivity of BVDV in the quadruple assay is slightly lower than that of the single qPCR method.Due to its sensitivity, specificity, simplicity, and rapidity, the RPA-CRISPR/Cas12a assay is more practical for on-site detection of cattle diarrhea pathogens than any existing detection method.
Management System for the Fattening Process of Bovines in Rotational Grazing using Diagnosis and Recommendation Systems
Rodrigo Garcia, Charles Benitez, Jose Aguilar
Cattle breeding has been one of the most important industrial sectors in the world since it is related to food security and the survival of the human race. Management of the cattle fattening process is a fundamental procedure for cattle breeders because it allows them to make strategic decisions, such as timely treatment in case of any abnormality (e.g., weight gain in herds, in their paddocks). This article aims to present a management system for the cattle fattening process under a rotational grazing scheme, considering the health status of the animal and the pasture, which should diagnose weight loss or gain in bovines and recommend actions when is required. The diagnostic process is based on a fuzzy system that defines rules that characterize the diagnostic process to determine the current situation given an input. Furthermore, the fuzzy classifier optimizes its rules by means of genetic algorithms by modifying its membership functions, providing a more accurate system for diagnosis. On the other hand, the recommendation system is based on a classification model of pasture crops, in which the best pasture is recommended given the soil variables. We tested our proposal with experimental cases, with promising results. For the fuzzy classifier, the accuracy metrics are very good, with values of accuracy close to 100% and of Area Under the Curve close to 1. For the classification model were used several machine learning techniques, resulting in the best classifier the random forest technique, with an accuracy of 98.61%.
Electronic computers. Computer science
Changes in biomarkers of metabolic stress during late gestation of dairy cows associated with colostrum volume and immunoglobulin content
Renato M. Rossi, Faith M. Cullens, Paola Bacigalupo
et al.
ABSTRACT: The objective of this observational study was to compare the metabolic status of dairy cows during the last 6 wk of gestation based on colostrum volume and immunoglobulin content. For this, healthy Holstein cows were randomly selected from 3 commercial herds in Michigan. In each farm, 4 cohorts of 21 cows (1 per season), stratified by parity, were enrolled (n = 228). Cows were blood sampled weekly during the last 6 wk of gestation, and biomarkers related to nutrient utilization, oxidant status, and inflammation were quantified in serum. Cows were milked within 6 h of calving, and the volume of colostrum produced was recorded and an aliquot collected. Concentration of IgG, IgA, and IgM were measured by radial immunodiffusion. Cows were grouped into high or low colostrum producer, high or low IgG, high or low IgA, and high or low IgM. For volume category, we arbitrarily defined 6 L of colostrum (4 L for first and 2 L for second feeding of calves) as the cutoff point, whereas for IgG we used the industry standard of ≥50 g/L. To create groups of low and high IgM or IgA, we used the median of these immunoglobulin as the cutoff point. Colostrum volume was lowest in winter, but no differences were observed among parity groups. Conversely, colostrum IgG concentration was highest in fall and winter, but colostrum IgM was lowest at these seasons. However, colostrum immunoglobulin content only showed a negative weak correlation with volume (Spearman's correlation coefficient < −0.28). Compared with low colostrum producer, high colostrum producer cows had higher concentrations of antioxidant potential and β-hydroxybutyrate, and lower cholesterol and oxidant status index. Cows with high IgG showed higher concentrations of glucose compared with low IgG. Cows with high IgA had higher concentrations of cholesterol, reactive oxygen and nitrogen species, oxidant status index, and total protein, whereas β-hydroxybutyrate and glucose were lower compared with low IgA. Biomarkers of metabolic stress were not significantly different between high IgM and low IgM. Nevertheless, the differences observed did not result in differences in inflammatory status between animals in any of the colostrum variable categories analyzed, suggesting that physiological homeostasis was not disrupted during late gestation in association with the colostrum variables studied. Overall, the great variability observed in colostrum variables suggests that colostrogenesis is a complex and multifactorial process. However, our results suggest that greater availability of antioxidants during late gestation could support the production of higher volumes of colostrum, which needs to be explored in future trials.
Dairy processing. Dairy products, Dairying
Camera traps reveal the natural corridors used by mammalian species in eastern Mexico
Jonathan O. Huerta-Rodríguez, Octavio C. Rosas-Rosas, Lauro López-Mata
et al.
Abstract Background Habitat loss and fragmentation in the Sierra Madre Oriental (SMO) ecological corridor have negative impacts on the movement and distribution of mammalian species that are of great ecological and evolutionary importance. Part of the SMO ecological corridor that is located in the state of Hidalgo is less studied despite being a potential dispersal route for mammals. The objectives of this study were to evaluate the presence and activity of terrestrial mammals in the riparian and non-riparian zones of SMO. Results Camera traps detected 15 mammalian species (i.e., 14 native and 1 domestic) in the non-riparian zone, and 12 mammalian species in the riparian zone (i.e., 10 wild and 2 domestic). The riparian corridor was mainly used by opportunistic medium-sized carnivores, while large carnivores and their potential prey were more frequent in the non-riparian zone. Conclusions Our findings suggest terrestrial mammals avoid the use of natural corridors due to the presence of domestic dogs, cattle and humans and look for new dispersal routes to move through their habitat and find the resources they need to survive. Even though some species can use disturbed corridors to move and find resources, they will change their activity patterns to avoid contact with humans and potential threats like dogs. It is, therefore, crucial to identify not one, but several corridors that must be preserved to improve the connectivity of terrestrial mammals in disturbed landscapes.
<i>Quo vadis</i> Cardiac Glycoside Research?
Jiří Bejček, Michal Jurášek, Vojtěch Spiwok
et al.
Cardiac glycosides (CGs), toxins well-known for numerous human and cattle poisoning, are natural compounds, the biosynthesis of which occurs in various plants and animals as a self-protective mechanism to prevent grazing and predation. Interestingly, some insect species can take advantage of the CG’s toxicity and by absorbing them, they are also protected from predation. The mechanism of action of CG’s toxicity is inhibition of Na<sup>+</sup>/K<sup>+</sup>-ATPase (the sodium-potassium pump, NKA), which disrupts the ionic homeostasis leading to elevated Ca<sup>2+</sup> concentration resulting in cell death. Thus, NKA serves as a molecular target for CGs (although it is not the only one) and even though CGs are toxic for humans and some animals, they can also be used as remedies for various diseases, such as cardiovascular ones, and possibly cancer. Although the anticancer mechanism of CGs has not been fully elucidated, yet, it is thought to be connected with the second role of NKA being a receptor that can induce several cell signaling cascades and even serve as a growth factor and, thus, inhibit cancer cell proliferation at low nontoxic concentrations. These growth inhibitory effects are often observed only in cancer cells, thereby, offering a possibility for CGs to be repositioned for cancer treatment serving not only as chemotherapeutic agents but also as immunogenic cell death triggers. Therefore, here, we report on CG’s chemical structures, production optimization, and biological activity with possible use in cancer therapy, as well as, discuss their antiviral potential which was discovered quite recently. Special attention has been devoted to digitoxin, digoxin, and ouabain.
Socioeconomic and socioecological issues of the pandemic crisis in the Amazon
Lise Tupiassu, Ana Elizabeth Neirão Reymão, Jean-Raphaël Gros-Désormeaux
The spread of covid-19 in the Amazon region urges us to reflect on how this can be related to forms of appropriation of the Amazon’s natural resources by global value chains. This article aims at discussing how the coronavirus pandemic has laid bare the vulnerabilities resulting from the Amazon’s insertion to global capitalism. Whether dependent on domestic or extraterritorial economic dynamics, no sector of activity and no community appears to be untouched by covid-19. The pandemic shows that revenues generated by economic activities such as mining and cattle breeding in the Amazon deeply depend on the international reach of their economic circuits, at the same time as on the non-observance of a number of fundamental rights of indigenous people. This clearly implies looking for alternatives to the unsustainable exploitation of nature and reconsidering the development model of the Amazon.
Prevalence of Gastrointestinal Protozoa on Bali Cattle in Lopok Sub-District, Sumbawa District
Aldi Hamdani, Nunuk Dyah Retno Lastuti, Yeni Dhamayanti
et al.
This study aims to determine the prevalence of gastrointestinal protozoa on Bali cattle in Lopok Sub-District, Sumbawa District. The study was conducted from January to April 2021 by taking 100 samples of Bali cattle feces from 7 villages in Lopok Sub-District, Sumbawa District. Samples were examined by sedimentation and floating methods. Species identification was determined by the morphology of the protozoa. A total of 62 (62%) samples were positively infected with protozoa. There were 4 species of gastrointestinal protozoa found infecting Bali cattle, namely Eimeria sp., Blastocytis sp., Entamoeba sp., and Balantidium sp. Most of the protozoa found in this study were Eimeria sp. (54%) and Blastocytis sp. (5%). This study found 59 samples (59%) were infected by one type of protozoa, 3 samples (3%) were infected by two types of protozoa. Cattle aged 0-6 months (81%) had a higher prevalence rate and were more susceptible to protozoal infections than cows aged 7 months – 2 years (70.3%) and more than 2 years (45.2%).
Infectious and parasitic diseases
Genetic variability of milk proteins in two cattle breeds of Piedmont region and the potential effects on milk quality
Melanie Salmasi, Riccardo Moretti, Stefania Chessa
et al.
In a scenario of climate change and risk of losing biodiversity, the safeguard of locally adapted breeds and their genetic reservoirs assumes a key role. Furthermore, these breeds are linked to their rearing territory maintenance also through the nutritional, economic or social value of their products. This study focussed on the analysis of milk protein genetic variability of the Piedmontese, the most abundant beef Italian breed, and the Barà-Pustertaler breed, through the testing of individual milk samples by isoelectrofocusing. Allelic and haplotypes frequencies were analysed and the presence of rare variants (CSN1S1*G, 0.7%, and CSN2*C, 0.7%, in Barà and Piedmontese breeds, respectively) was detected. The analysed breeds showed allelic frequencies close to the one found in breeds with good cheesemaking aptitude, like the Italian Brown and Reggiana breeds (e.g. CSN2*B, 20.7% and 16.4%, and CSN3*B, 41,1% and 35%, in Piedmontese and Barà breeds, respectively), and are a good reservoir of alleles that are decreasing (e.g. CSN2*B) in Italian Holstein. Moreover, they have higher frequencies of interesting haplotypes (e.g. BA1B, 8% and 11.2%, and BA2B, 20.8 and 13.1% in Piedmontese and Barà breeds, respectively) that could positively influence composition, nutritional, and technological properties of milk. These breeds could be used to further investigate the effects of variants and haplotypes that could help support the evolution and the changing demands of the dairy sector.Highlights In order to address the problem of climate change effects on animal biodiversity, local breeds should be safeguarded and valorised. The analysed breeds have discrete frequencies of milk protein variants and casein haplotypes associated with good technological properties. Knowing the genetic background of non-dairy commercial breeds is the first step to preserve potentially useful genetic variability.
Equestrian culture and heritage in Brazil: untapped potential for a tourism that favours local development? São Luiz do Purunã as a case study
Miriam Adelman
Brazil is one of the world’s “horse giants”, home to the world’s fourth largest equine population, and to a popular equestrian culture that has yet to be duly recognized as heritage. This culture, emerging from its historic link to cattle ranching and agriculture, is also reshaped by currently expanding sport and leisure uses of the horse. With a focus on the needs and predicaments of rural communities in which horses are an important symbolic and cultural resource – perhaps more than an economic one – I provide a description and discussion of my ethnographic research within the village of São Luiz do Purunã, located in a region of environmental protection in southern Brazil. As a case study that examines the different social actors involved in turistification processes, my research contributes to debates on the patrimonialisation of traditional equestrian culture, its promise and its contradictions.
SEROLOGICAL AND MOLECULAR DETECTION OF COWPAE MOSAIC VIRUS INFECTING COWPAE IN IRAQ
Abd Al-Hakeem A. Al-Shamary, Nawres A. Al-Kuwaiti
This study was conducted to detect CPMV in infecting cowpea plantes at Plant Protection Department/College of Agriculture, University of Baghdad. Symptomatic cowpea plants collected from fields in Baghdad and Diyala Provinces were tested by Emzyme-linked immunosorbent assay (ELISA), reverse transcription- polymerase chain reaction RT-PCR and Immuno-capture reverse transcription-polymerase chain reaction (IC-RT-PCR) using commercial kits. ELISA approach could detect Cowpea mosaic virus CpMV in tested samples but with virus concentration lower than the positive control supplied with the kit, indicating a unique virus strain. RNA from samples showed the highest CpMV concentration were extracted by direct release and used in ICPT-PCR using como2 a comovirus specific primer set. cDNA was synthetized using como2 revers primer. RT-PCR confirmed the detection of comoviruses when amplified 200 bp DNA fragments from CpMV infected samples. Sequence analysis confirmed CpMV detection when compared with equivalent GenBank sequences. The sequence obtained shared 94.1% maximum nucleotide identity with RNA dependent RNA polymerase (RdRp) gene of CpMV from Egypt (Acc. No. KT438619 and X00206). Both sequence analysis and IC-RTPCR indicated that virus isolate detected could be an Iraq divergent strain of CpMV.
Agriculture (General), Forestry
ENTOMOLOGICAL ASPECTS OF LUMPY SKIN DISEASE EPIZOOTOLOGY (REVIEW)
Ya. E. Pestova, A. V. Kononov, A. V. Sprygin
Lumpy skin disease (LSD) is a serious threat to the global cattle farming, including that in the Russian Federation where the frst outbreak was reported in 2015. Since the disease occurred for the frst time, it has continued to spread in this country; however, virus transmission mechanisms have not yet been studied. Transmission through insect bites is considered to be the most likely mechanism of virus shortrange transmission. At present, such arthropod species as stable fly (Stomoxys сalcitrans), Aedes aegypti mosquitoes, as well as Amblyomma hebraeum and Rhipicephalus appendiculatus ticks are regarded as potential vectors. Viral DNA has also been detected on the exoskeletons of house flies (Musca domestica). The available literature describes the results of many studies on the role of arthropods in LSD virus spread, but the data presented are inconsistent and do not provide an unambiguous answer concerning the level of signifcance of potential LSD virus vectors in the progression of the feld epizootic. These papers investigate the ability of gadflies, flies and ticks to act as mechanical vectors. Currently, there is no unequivocal viewpoint with respect to the proved LSD vector. This paper reviews the entomological papers aimed at studying possible LSD virus transmission by arthropods.
Replacing sodium bicarbonate with half amount of calcareous marine algae in the diet of beef cattle
Carlo Angelo Sgoifo Rossi, Riccardo Compiani, Gianluca Baldi
et al.
ABSTRACT This study evaluated the effects of feeding calcareous marine algae or sodium bicarbonate as rumen buffer on the performance, behaviour, in vitro diet digestibility, and meat quality of beef cattle. A total of 180 Charolaise bullocks (536±38 kg; 14±1 months of age) were divided into two homogeneous groups and fed a diet with a mineral mix containing 40% sodium bicarbonate or 20% calcareous marine algae (CMA) for the entire fattening period (130 days). Of the in vivo and in vitro parameters evaluated, CMA supplementation improved average daily gain and feed conversion ratio and reduced the prevalence of bloat and lameness. Bullocks fed CMA tended to exhibit a calmer behaviour while in the pen. Supplementation with CMA improved rumen pH and in vitro digestion. Meat from bullocks fed CMA showed a lower pH and higher lightness and tenderness. The results suggest that CMA is more effective than sodium bicarbonate in buffering beef cattle, with a positive impact on growth performance, feed efficiency, health, and meat quality.
Correction: Molecular epidemiology and risk factors of Anaplasma spp., Babesia spp. and Theileria spp. infection in cattle in Chongqing, China.
PLOS ONE Staff
[This corrects the article DOI: 10.1371/journal.pone.0215585.].
Evaluation of 17 microsatellite markers for parentage testing and individual identification of domestic yak (Bos grunniens)
Jie Pei, Pengjia Bao, Min Chu
et al.
Background Yak (Bos grunniens) is the most important domestic animal for people living at high altitudes. Yak ordinarily feed by grazing, and this behavior impacts the accuracy of the pedigree record because it is difficult to control mating in grazing yak. This study aimed to evaluate the pedigree system and individual identification in polled yak. Methods A total of 71 microsatellite loci were selected from the literature, mostly from the studies on cattle. A total of 35 microsatellite loci generated excellent PCR results and were evaluated for the parentage testing and individual identification of 236 unrelated polled yaks. A total of 17 of these 35 microsatellite loci had polymorphic information content (PIC) values greater than 0.5, and these loci were in Hardy–Weinberg equilibrium without linkage disequilibrium. Results Using multiplex PCR, capillary electrophoresis, and genotyping, very high exclusion probabilities were obtained for the combined core set of 17 loci. The exclusion probability (PE) for one candidate parent when the genotype of the other parent is not known was 0.99718116. PE for one candidate parent when the genotype of the other parent is known was 0.99997381. PE for a known candidate parent pair was 0.99999998. The combined PEI (PE for identity of two unrelated individuals) and PESI (PE for identity of two siblings) were >0.99999999 and 0.99999899, respectively. These findings indicated that the combination of 17 microsatellite markers could be useful for efficient and reliable parentage testing and individual identification in polled yak. Discussion Many microsatellite loci have been investigated for cattle paternity testing. Nevertheless, these loci cannot be directly applied to yak identification because the two bovid species have different genomic sequences and organization. A total of 17 loci were selected from 71 microsatellite loci based on efficient amplification, unambiguous genotyping, and high PIC values for polled yaks, and were suitable for parentage analysis in polled yak populations.
Medicine, Biology (General)
The Studies on In Vivo Embryo Production using Sex-Sorted Sperm in Korean Brindle Cattle
Hyang-A Jeon, Seung-Ju Moon, Ji-Woong Lee
et al.
The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be 6.20±2.28/donor from the control group and was significantly lowly recovered to be 1.57±1.72/donor from the group treated at a sperm concentration of 10×106 (p<0.05). The number of unfertilized embryo was 0.8±1.30/donor in control group which was significantly lower than the group treated at a sperm concentration of 4×106 (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.
Biotechnology, Medicine (General)
Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos
D.K. de Souza, L.P. Salles, A.A.M. Rosa e Silva
Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.
Medicine (General), Biology (General)
Physiological level production of antigen-specific human immunoglobulin in cloned transchromosomic cattle.
Akiko Sano, Hiroaki Matsushita, Hua Wu
et al.
Therapeutic human polyclonal antibodies (hpAbs) derived from pooled plasma from human donors are Food and Drug Administration approved biologics used in the treatment of a variety of human diseases. Powered by the natural diversity of immune response, hpAbs are effective in treating diseases caused by complex or quickly-evolving antigens such as viruses. We previously showed that transchromosomic (Tc) cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin heavy-chain (hIGH) and kappa-chain (hIGK) germline loci (named as κHAC) are capable of producing functional hpAbs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, are homozygously inactivated (double knockouts or DKO). However, B lymphocyte development in these Tc cattle is compromised, and the overall production of hpAbs is low. Here, we report the construction of an improved HAC, designated as cKSL-HACΔ, by incorporating all of the human immunoglobulin germline loci into the HAC. Furthermore, for avoiding the possible human-bovine interspecies incompatibility between the human immunoglobulin mu chain protein (hIgM) and bovine transmembrane α and β immunoglobulins (bIgα and bIgβ) in the pre-B cell receptor (pre-BCR) complex, we partially replaced (bovinized) the hIgM constant domain with the counterpart of bovine IgM (bIgM) that is involved in the interaction between bIgM and bIgα/Igβ; human IgM bovinization would also improve the functionality of hIgM in supporting B cell activation and proliferation. We also report the successful production of DKO Tc cattle carrying the cKSL-HACΔ (cKSL-HACΔ/DKO), the dramatic improvement of B cell development in these cattle and the high level production of hpAbs (as measured for the human IgG isotype) in the plasma. We further demonstrate that, upon immunization by tumor immunogens, high titer tumor immunogen-specific human IgG (hIgG) can be produced from such Tc cattle.
Induction of Pulmonary Edema and Emphysema in Cattle and Goats with 3-Methylindole
J. Carlson, M. Yokoyama, E. O. Dickinson
Dairy Cattle
G. J. Gallivan, W. McDonell
Variations between and within breeds of cattle in resistance to field infestations of the cattle tick (Boophilus microplus).
G. Seifert