Calves In Japan Research Articles

A calf with hind limb paralysis and dysstasia and a genome sequence analysis of an isolated Clostridium perfringens toxinotype E strain.

Clostridium perfringens toxinotype E infections are rare in calves, and the development of intestinal lesions were commonly observed. In 2012, a 6-day-old calf in Japan exhibited swelling with emphysema on the right gluteal region, sudden paralysis of the hind limb and dysstasia. A pathological examination revealed myositis of the gluteal muscle and neuritis of the ischiatic nerve. C. perfringens type E strain CP118 was isolated from the affected muscle. However, the intestinal symptoms and lesions that commonly develop in type E infections in calves were not detected in the present case. Genome analyses revealed that CP118 possessed 16 virulence-related genes, including enterotoxin, and was closely related to other type E and F strains. Particularly, CP118 was more closely related to type E strains from humans, including a food poisoning case, than calf isolates, suggesting its potential to cause food poisoning in humans and, thus, its importance as a potential risk to public health. Since CP118 did not possess the reported toxin genes associated with neuropathy, pyogenic inflammation caused by CP118 and/or other bacteria may have damaged the ischiatic nerve, resulting in neuropathy. Alternatively, unidentified CP118 toxins may have caused the neuropathy. This is the first study to report C. perfringens type E infection with peripheral neuropathy. The distribution of all the reported virulence-related genes in the C. perfringens population as well as the details of this rare case will provide further insights into C. perfringens type E infections.

The first genetic characterization of Setaria marshalli (Nematoda, Spirurida) with reliable DNA barcoding based on a mitochondrial genetic marker.

Setaria marshalli is a mosquito-borne filarial nematode that causes infection in calves younger than two years old. In the present study, nematodes were obtained from a calf in Japan and morphologically identified as S. marshalli. Additionally, the partial cytochrome oxidase subunit I (COI) region (596 bp) was analyzed for the first time to establish a reliable DNA barcode. Nucleotide sequences of COI were identical among the seven worms obtained. The COI region can be a useful marker for species discrimination in the case of S. marshalli since nucleotide variations observed between the closest congener, Setaria cervi (51/596 bp), were sufficient to allow species discrimination. However, the phylogenetic relationship of S. marshalli with its congeners was unclear in a maximum likelihood tree. We found that the partial COI sequence of S. marshalli analyzed in the present study matched a relevant section of the complete mitochondrial genome of S. labiatopapillosa that was deposited in the International Nucleotide Sequence Database. This finding suggests that S. marshalli was misdiagnosed as S. labiatopapillosa in a previous study. It is crucial to conduct accurate morphological analyses to obtain reliable molecular information regarding Setaria nematodes.

Distribution of Cryptosporidium species isolated from diarrhoeic calves in Japan.

Cryptosporidium spp. are enteric protozoan parasites that infect a wide range of hosts including humans, and domestic and wild animals. The aim of this study was to molecularly characterize the Cryptosporidium spp. found in calf faeces in Japan. A total of 80 pre-weaned beef and dairy calves' diarrhoeic faecal specimens were collected from nine different prefectures in Japan. A nested polymerase chain reaction targeting the small subunit 18S rRNA and GP60 genes were used to detect the Cryptosporidium genotypes and subtypes. 83.8% (67 out of 80) of the specimens were positive for Cryptosporidium spp.; Cryptosporidium was found in both beef and dairy calves. Cryptosporidium parvum was the predominant species, detected in 77.5% (31/40) of beef calves and 80% (32/40) of dairy calves. Cryptosporidium bovis was also detected, 5.0% (2/40) of dairy calves, and C. ryanae was also found 2.5% (1/40) of dairy calves. One mixed-species infection, 2.5% (1/40) was detected in a beef calf having C. parvum, and C. ryanae. We detected the most common subtype of C. parvum (i.e., IIaA15G2R1), as well as other subtypes (i.e., IIaA14G3R1, IIaA14G2R1, and IIaA13G1R1) that have not previously been detected in calves in Japan. Our results demonstrate the widespread diversity of Cryptosporidium infection in calves in Japan.

Outbreak of sudden death in weaned calves on a New York dairy caused by Strongyloides papillosus

S. papillosus are slender hair-like worms that have a predilection for the duodenum and proximal jejunum of ruminants. The third larval stage of S. papillosus penetrates the skin, often in the coronary band region of the limb, and migrates to the gastrointestinal tract where it matures to the parthenogenetic adult stage. Typically S. papillosus infection of calves in North America is associated with clinical signs of diarrhea, dehydration, anorexia, and weight loss or poor weight gain (Thamsborg et al, 2017).
 Outbreaks of sudden death associated with heavy S. papillosus burden are rare and have only been reported in 2-5 month old calves in Japan (Taira and Ura, 1991). These Japanese outbreaks occurred in the summer, where common conditions included sawdust bedding, high environmental temperatures, and high humidity. Experimental studies revealed the sudden death of calves with heavy worm burdens to be preceded by cardiac arrhythmias for 1-2 days and ventricular arrhythmias leading to cardiac arrest (Tsuji et al, 1992). These researchers hypothesized that S. papillosus may produce a cardiotoxin that leads to cardiac arrest of infected calves.
 S. papillosus has recently been implicated as the novel cause of an outbreak of sudden death in weaned dairy calves on a well-managed farm in New York State. The following report describes this first documented North American outbreak of sudden death in calves caused by S. papillosus.

Mycoplasma bovis isolates from dairy calves in Japan have less susceptibility than a reference strain to all approved macrolides associated with a point mutation (G748A) combined with multiple species-specific nucleotide alterations in 23S rRNA.

Erythromycin, tylosin and tilmicosin are approved for use in cattle in Japan, the latter two being used to treat Mycoplasma bovis infection. In this study, 58 M. bovis isolates obtained from Japanese dairy calves all exhibited reduced susceptibility to these macrolides, this widespread reduced susceptibility being attributable to a few dominant lineages. All 58 isolates contained the G748A variant in both the rrl3 and rrl4 alleles of 23S rRNA, whereas a reference strain (PG45) did not. G748 localizes in the central loop of domain II (from C744 to A753) of 23S rRNA, which participates in binding to mycinose, a sugar residue present in both tylosin and tilmicosin. A number of in vitro-selected mutants derived from M. bovis PG45 showed reduced susceptibility to tylosin and tilmicosin and contained a nucleotide insertion within the central loop of domain II of rrl3 (U747-G748Ins_CU/GU or A743-U744Ins_UA), suggesting that mutations around G748 confer this reduced susceptibility phenotype. However, other Mycoplasma species containing G748A were susceptible to tylosin and tilmicosin. Sequence comparison with Escherichia coli revealed that M. bovis PG45 and isolates harbored five nucleotide alterations (U744C, G745A, U746C, A752C and A753G) in the central loop of domain II of 23S rRNA, whereas other Mycoplasma species lacked at least two of these five nucleotide alterations. It was therefore concluded that G748 mutations in combination with species-specific nucleotide alterations in the central loop of domain II of 23S rRNA are likely sufficient to reduce susceptibility of M. bovis to tylosin and tilmicosin.

Effects of temperature-humidity index on health and growth performance in Japanese black calves.

We evaluated the relationship between the temperature-humidity index (THI) and health and growth performance in Japanese black calves in Japan. Data were collected from medical records of 19,313 Japanese black calves aged up to 3months for correlation analysis with THI from July 2008 to June 2011. Data were also collected on the market weights of 57,144 Japanese black calves, and we calculated the body weight gain (BWG) of each calf based on body weight and age in days at the calf market. Analysis for the relationship between disease incidence and THI demonstrated a negative correlation (r = -0.54, p < 0.01). In addition, the mean disease incidence at THI of ≤50 was significantly higher than that at THI of ≥71. Analysis for the relationship between growth performance in calves and THI revealed that a lower THI during the month of birth was associated with a lower BWG at the calf market and that BWG with THI of ≤70 was significantly lower than that with THI of ≥71 (p < 0.05). In contrast, a higher mean THI during the third month after birth was associated with a lower BWG at the calf market, and BWG with THI of >75 was significantly lower than that with THI of ≤50 or THI ranging from 56 to 60 (p < 0.05). These results indicate that Japanese black calves are susceptible to a cold environment immediately after birth, whereas they are susceptible to a heat environment 3months after birth.

Detection of avian-like rotavirus A VP4 from a calf in Japan.

A total of 568 normal feces from calves on a beef farm in Fukui Prefecture, Japan, in 2011–2012 were examined by RT-semi-nested PCR for rotavirus A (RVA) VP4 genes. Through partial sequencing and BLAST analyses of 84 VP4-positive specimens, we identified an avian-like RVA strain, N2342, which shares highest nucleotide identity (80.0%) with known avian-like bovine strain 993/83, in one specimen. Phylogenetic analysis also revealed a close genetic relationship between N2342 and avian RVAs, suggesting bird-to-cattle transmission. We observed frequent contact of wild birds with calves in the farm, suggesting that these birds were the source of the virus.

Nonsuppurative Encephalomyelitis in a Calf in Japan and Isolation of Japanese Encephalitis Virus Genotype 1 from the Affected Calf

Japanese encephalitis virus (JEV) was isolated from the cerebrum of a calf which showed severe neurological symptoms in late September 2009, and the JEV isolate was revealed to be of genotype 1. This is the first report describing the isolation of genotype 1 JEV from cattle.

Molecular Characterization of &lt;i&gt;Cryptosporidium&lt;/i&gt; Isolates from Calves in Ishikari District, Hokkaido, Japan

Cattle are major hosts of Cryptosporidium. Cryptosporidiosis in neonatal calves is associated with retarded growth, weight loss and calf mortality, and zoonotic infections in humans. Fecal samples were collected from calves in Ishikari District, Hokkaido, Japan and examined by PCR and sequence analyses. Among the 107 fecal samples collected in May and June 2012, 25 (23%) were positive for Cryptosporidium, including 8 samples (7%) having C. parvum, 10 (9%) having C. bovis and 7 (7%) having C. ryanae. This is first time C. ryanae has been detected in Hokkaido. Furthermore, it is the first detection of C. ryanae from pre-weaned calves in Japan. Microscopic observation with the flotation method is powerful and traditional tool for screening for Cryptosporidium species, but it sometimes leads to low detection of Cryptosporidium with low oocyst shedding intensity. If calves with or without diarrhea are examined using the molecular diagnostic tools, C. bovis and C. ryanae might be detected in other areas of Japan including Hokkaido. Here, the zoonotic species, C. parvum, was also observed. Therefore, calves can be potential sources of cryptosporidial infections for humans and other animals. The detection of C. parvum was statistically correlated with diarrhea in calves.

Molecular Analysis of Parapoxvirus Detected in Eight Calves in Japan

ABSTRACTMolecular analysis of parapoxvirus envelope genes was performed. Parapoxvirus DNA was detected in eight calves from eight farms in Iwate Prefecture, Japan, between April and September 2010. Seven of the detected viruses were identified as bovine papular stomatitis virus (BPSV) by sequencing, because their nucleotide identity was more than 96.8% similar compared with BPSV strain V660. Among them, two formed a subgroup, because their amplicons were digested with Xmn I (a marker for BPSV) and Hinc II and exhibited a T61C nucleotide substitution in the sequenced region. The remaining virus was pseudocowpox virus that had not been reported previously in Japan. Our results demonstrate the presence of a new BPSV variant in Japan with genetic variability in the envelope gene.

Molecular epidemiology of rotaviruses among healthy calves in Japan: Isolation of a novel bovine rotavirus bearing new P and G genotypes

A total of 171 fecal specimens collected from healthy calves on a beef farm in Gifu Prefecture, Japan in 2006–2007 were examined for group A rotaviruses by RT-semi-nested PCR targeting the coding region for VP8*. Nine specimens were positive for rotavirus. G and P genotyping indicated that one strain was G10P[11]-like and six strains were considered to be the same unknown G and P genotypes. Among these six untypeable strains, one strain, AzuK-1, was adapted to cell culture and analyzed. Sequence and phylogenetic analyses of the full lengths of VP4 and VP7 genes revealed that AzuK-1 strain is a novel bovine rotavirus bearing new G21 and P[29] genotypes as confirmed by the RCWG. Furthermore, we detected G21P[29] rotaviruses in fecal specimens collected from healthy calves in Hokkaido, Japan during the period from 1997 to 1998. These findings suggest that novel G21P[29] rotaviruses have been widely prevalent among cattle for over 10 years in Japan.

Identification of Genotypes of Giardia intestinalis Isolates from a Human and Calf in Japan

Giardia intestinalis is recognized as a significant pathogen in humans and animals, causing diarrhea. Recent molecular studies indicate that G. intestinalis is composed of genetically distinct multiple genotypes. Therefore, it is valuable to distinguish among genotypes in the epidemiology of Giardia infection in humans and animals. Although G. intestinalis has been found in humans and animals in Japan, the genotype of isolates remains unclear except for several isolates from dogs, because identification has been performed only by conventional microscopy. We report herein the genotypes of G. intestinalis isolates distinguished by a phylogenetic analysis. G. intestinalis isolates originated from a patient and a calf were found to have Assemblage B and E, respectively.

Genetic parameters for packed red blood cell volume and daily gain during grazing in Japanese Shorthorn calves facing a natural challenge from Japanese theileriosis

Japanese theileriosis, which is caused by the intraerythrocytic protozoan Theileria orientalis sergenti, is one of the most harmful anemic diseases to grazing calves in Japan. In this paper, we estimated genetic and phenotypic parameters of packed cell volume (PCV) and productivity in 435 Japanese Shorthorn calves, and discussed the feasibility of the genetic improvement for higher anemia tolerance. The heritabilities of PCV at pre-grazing (PrePCV) and minimum PCV during grazing period (MinPCV), age at MinPCV (MinAge), the times of theileriacidal drug treatment (TRT) and daily body weight gain during the grazing period (DG) were 0.54±0.11, 0.13±0.06, 0.21±0.07, 0.35±0.21 and 0.65±0.06, respectively. There were weakly negative genetic correlations between MinPCV and DG. MinAge had a strongly positive genetic correlation with MinPCV, while a strongly negative genetic correlation with DG. TRT had negative phenotypic and genetic correlations with all traits. From the results of simulations of selection, a single improvement in DG would reduce MinPCV and TRT in a population as a correlated response. Although it would lead to a reduction in TRT as a correlated response, it would also lead to temporarily severe anemia. This would require more labor for management of grazing herds and frequent times of sanitary check. On the other hand, a simultaneous improvement in DG and anemia tolerance would be inferior in efficiency of improvement in DG. Instead of a low efficiency of improvement in DG, a simultaneous improvement would decrease TRT more efficiently and increase MinPCV.

Prevalence of Cryptosporidium parvum infection and pattern of oocyst shedding in calves in Japan

Cryptosporidium parvum infection and the pattern of oocyst shedding were observed in calves. A total of 480 fecal samples were collected from 30 calves (age, ≤ 30 days) over a period of 10 months from June 1998 to March 1999. A sucrose centrifugal flotation technique revealed 28/30 (93%) calves were passing Cryptosporidium oocysts. Oocyst shedding was first detected on the sixth day after birth, with 8% of the calves testing positive. This rate increased day by day and reached approximately 80% by day 15. Oocyst shedding varied from 1 to 13 days, with a mean of 7 days. Calves infected with C. parvum had a significantly higher rate of diarrhea (33%) than non-infected calves (8%) ( P<0.05), suggesting C. parvum infection as the likely cause. The mean number of oocysts excreted by calves ≤ 30 days old was approximately 6×10 7 per gram of feces. These results indicated that one calf would excrete some 6×10 11 oocysts in the first month after birth, taking both the quantity of feces in a day and the period of excretion into consideration. Accordingly, it is clear that calves are important in the spread of cryptosporidiosis to calves and humans.

Control of Theileria sergenti infection by vaccination.

Bovine piroplasmosis caused by Theileria sergenti is a major cause of economic loss in grazing cattle in Japan. Infected calves show chronic anaemia with intraerythrocytic piroplasms and occasionally die in severe cases. We found that parasite stocks and isolates consist of genetically and antigenically mixed populations. To differentiate parasite populations bearing 3 allelic forms of p32/34, an immunodominant piroplasm surface protein, 3 sets of oligonucleotide primers were designed to amplify either of 3 alleles by polymerase chain reaction (PCR). By using this allele-specific PCR, we found that the majority of T. sergenti-infected calves in Japan harbored mixed parasite populations bearing C and I type parasites. To control Theileria infection, we produced 2 vaccine candidates: recombinant baculovirus p32 and synthetic peptide containing Lys-Glu-Lys (KEK) motif. Immunization with either recombinant p32 or synthetic peptide containing KEK sequences with Freund's complete adjuvant resulted in low parasitemia and reduced the clinical symptoms compared to control calves. Interestingly, the parasite with the p32 allelic form corresponding to the one used as the immunogen was suppressed.

Use of the Bo-RBC-SCID Mouse Model for Isolation of a Babesia Parasite from Grazing Calves in Japan

SCID mice with circulating bovine red blood cells (Bo-RBC-SCID mice) were used to isolate Babesia parasites from grazing calves in Japan. Although the initial blood samples obtained from the calves contained both Babesia and Theileria parasites, we were able to isolate only the Babesia parasites by repeated blood passages in Bo-RBC-SCID mice, taking advantage of the much more rapid growth of Babesia than of Theileria. The parasites isolated had characteristics of large-type Babesia, showing typical paired pyriform morphology. The Bo-RBC-SCID mice infected with the Babesia parasites developed a high level of parasitemia, showing severe clinical symptoms characterized by hemoglobinuria, jaundice, and hemolytic anemia. In addition, some mice exhibited nervous symptoms, particularly paralysis of both posterior limbs. The results demonstrated that the Bo-RBC-SCID mouse model was useful not only for isolating Babesia from cattle but also for studying the disease caused by the Babesia infection.

Relative frequencies of G (VP7) and P (VP4) serotypes determined by polymerase chain reaction assays among Japanese bovine rotaviruses isolated in cell culture.

The relative frequencies of both the G (VP7) and P (VP4) serotypes of 40 bovine rotaviruses isolated in cell culture from diarrheic calves in Japan between January 1983 and February 1991 were determined by recently developed polymerase chain reaction assays. Isolates with G serotype 6 and P serotype 5 (UK-like strains) were most frequently found (42.5%) followed by isolates with G6P11 (17.5%), G6P1 (10%), or G10P5 (10%). Isolates with G10P11 (B223-like strains) were least frequently found (7.5%). The presence of various combinations of G and P serotypes suggests frequent reassortment in nature among bovine rotaviruses.

Selenium and Tocopherol Levels in Calves With White Muscle Disease of Cardiac and Skeletal Muscle Type in Japan

White muscle disease (WMD) in calves is a nutritional degeneration of skeletal and cardiac muscle caused by tocopherol deficiency1, selenium deficiency2 or both3. Author et al. researched WMD in livestock (calves, lambs, foals) in Japan and clarified that the disease is caused by deficiencies of selenium and tocopherol4,5,6). In this study, cases of WMD in calves in Japan were divided into skeletal muscle type with degeneration of skeletal muscle and cardiac muscle type with degeneration of cardiac muscle. We compared the clinico-pathological observations and the levels of tocopherol and selenium.

Hydranencephaly-cerebellar hypoplasia in a newborn calf after infection of its dam with Chuzan virus.

Chuzan virus at 2 to 3 passage levels in cell cultures after isolation was inoculated intravenously into 15 seronegative pregnant cows at 89 to 150 days of gestation. All of the cows developed viremia a few days after inoculation and antibodies 2 weeks after inoculation. No clinical signs, except leukopenia, were observed throughout the experimental period. These 15 cows delivered 15 calves after normal gestation. One of the calves which was born to a dam inoculated at 120 days of gestation, showed impairment of movement, and the remaining 14 were healthy. Postmortem examination revealed that this calf had hydranencephaly- cerebellar hypoplasia (HCH) syndrome and that the remaining calves were normal. Two of the 15 calves, including the one that had HCH syndrome, had antibody to Chuzan virus in their precolostral sera. These findings provide additional evidence that Chuzan virus is the etiological agent of an epizootic of congenital abnormalities with HCH syndrome of calves in Japan, 1985 to 1986. We propose to name the HCH syndrome caused by Chuzan virus infection Chuzan disease.

Kawanabe virus, an isolate from a calf in Japan: a new virus belonging to the New Jersey serotype of the epizootic hemorrhagic disease serogroup of genus Orbivirus.

[in Japanese]