Eficacia protectora de una vacuna inactivada contra el virus de la enfermedad hemorrágica del conejo tipo 2 preparada a partir de un aislado local en Egipto / Protective efficacy of an inactivated vaccine against Rabbit hemorrhagic disease virus 2 prepared from a local isolate in Egypt

Rabbit hemorrhagic disease is a contagious viral disease of rabbits controlled by vaccination. The present study was aimed to diagnose rabbit hemorrhagic disease from 11 infected farms from Qalubia governorate during 2019 and to prepare homologous vaccine against rabbit hemorrhagic disease virus 2. For this purpose, 11 liver samples were collected from suspected cases and subjected to detection and identification of circulating rabbit hemorrhagic disease virus. Ten samples were confirmed to be rabbit hemorrhagic disease virus using hemagglutination test, animal inoculation and reverse transcriptase polymerase chain reaction. Sequencing and phylogenetic analysis of two isolates (R5&R6) revealed the presence of rabbit hemorrhagic disease virus 2 (A/Qalubia/2019 and B/Qalubia/2019) under accession number MT07629 and MT067630 respectively. The inactivated rabbit hemorrhagic disease virus vaccines were prepared using Montanide ISA 206 oil or aluminum hydroxide gel adjuvants. Prepared vaccines were inoculated subcutaneously in susceptible rabbits and submitted to sterility, safety and potency tests. Obtained results showed that mean hemagglutination inhibition titer for aluminum hydroxide gel vaccine was 6,7.7,8.9 and 9.1 log2 while, Montanide vaccine reached to 6.7,8.7,9.2 and 9.5 log2 at 1st, 2nd, 3rd, and 4th weeks post vaccination, respectively. Immunized rabbits with Montanide vaccine showed better protection reach to 70 percent, 90 percent percent, 100 percent and 100 percent when compared to aluminum hydroxide gel vaccine 60 percent, 70 percent, 90 percent and 90 percent at 1st, 2nd, 3rd and 4th weeks post vaccination respectively. It was concluded that newly emerged rabbit hemorrhagic disease virus 2 was isolated from suspected cases. The two prepared vaccines were sterile, safe and potent. The oily adjuvanted rabbit hemorrhagic disease virus 2 vaccine stimulated an earlier and higher humoral immune response than the aluminum hydroxide gel adjuvanted vaccine. This humoral immune response achieved significant level of protection(AU)

La enfermedad hemorrágica del conejo es una enfermedad viral contagiosa de los conejos que se controla mediante vacunación. El presente estudio tuvo como objetivo diagnosticar la enfermedad hemorrágica del conejo en 11 granjas infectadas de la provincia de Qalubia, durante 2019 y preparar una vacuna homóloga contra el virus de la enfermedad hemorrágica del conejo tipo 2. Para este propósito, se recolectaron 11 muestras de hígado de casos sospechosos y se sometieron a detección e identificación de virus circulante de la enfermedad hemorrágica del conejo. Se confirmó que diez muestras eran positivas al virus de la enfermedad hemorrágica del conejo, utilizando para ello la prueba de hemaglutinación, inoculación en animales y Reacción en cadena de la polimerasa con transcriptasa inversa. La secuenciación y el análisis filogenético de dos aislamientos (R5 y R6) revelaron la presencia del virus de la enfermedad hemorrágica del conejo tipo 2 (A/Qalubia/2019 y B/Qalubia/2019) con los números de acceso MT07629 y MT067630 respectivamente. Las vacunas inactivadas del virus de la enfermedad hemorrágica del conejo se prepararon usando adyuvantes de gel de hidróxido de aluminio o aceite Montanide ISA 206. Las vacunas preparadas se inocularon por vía subcutánea en conejos susceptibles y se sometieron a pruebas de esterilidad, seguridad y potencia. Los resultados obtenidos mostraron que el título medio de inhibición de la hemaglutinación para la vacuna en gel de hidróxido de aluminio fue de 6; 7,7; 8,9 y 9,1 log2, mientras que la vacuna de Montanide alcanzó 6,7; 8,7; 9,2 y 9,5 log2 en la 1ª, 2ª, 3ª y 4ª semanas después de la vacunación, respectivamente. Los conejos inmunizados con la vacuna Montanide tuvieron una mejor protección, alcanzándose niveles de 70 por ciento, 90 por ciento, 100 por ciento y 100 por ciento en comparación con la vacuna en gel de hidróxido de aluminio 60 por ciento, 70 por ciento, 90 por ciento y 90 por ciento en la 1ª, 2ª, 3ª y 4ª semanas después de la vacunación, respectivamente. Se concluyó que el virus de la enfermedad hemorrágica del conejo tipo 2 de reciente aparición se aisló de los casos sospechosos. Las dos vacunas preparadas fueron estériles, seguras y potentes. La vacuna contra el virus de la enfermedad hemorrágica del conejo tipo 2 con adyuvante oleoso estimuló una respuesta inmune humoral más temprana y mayor que la vacuna con adyuvante en gel de hidróxido de aluminio. Esta respuesta inmune humoral confirió un nivel significativo de protección(AU)

Tandem expression of the major epitope domains of the Moschus chrysogaster hemorrhagic disease virus VP60 and its protective efficacy to rabbits / 生物工程学报

Moschus chrysogaster (sifanicus) viral hemorrhagic disease (McVHD) is an acute and highly lethal infectious disease caused by Moschus chrysogaster hemorrhagic disease virus (McHDV) whose genome sequence is highly homologous with rabbit hemorrhagic disease virus. To screen the protective antigen of McHDV and set the basis for study of McVHD vaccine, the antigen epitope of major structural protein VP60 of McHDV was analyzed, and the specific primers were designed to obtain three amplified DNA sequences encoding the main antigen epitope of VP60 from McHDV by using RT-PCR. Then the three DNA fragments were sequenced and cloned to prokaryotic expression vector with pET-28a(+) by using overlap extension PCR, and finally the prokaryotic expression plasmid pET-truncated-VP60 was constructed. Subsequently, the pET-truncated-VP60 was transformed into Escherichia coli BL21(DE3), and the recombinant proteins were expressed by IPTG induction. Finally, the expressed protein was purified and applied to immunize that without immunizing with RHD vaccine, then the antiserum titers were evaluated by the hemagglutination inhibition test, and the immune-protective efficacy of the recombinant proteins was observed and analyzed through animal challenge test. The results showed that the multi-epitope DNA fragments of VP60 of McHDV was successfully expressed in the form of inclusion bodies in E. coli, and the relative molecular weight of recombinant proteins is about 45 kDa. After immunized with the recombinant proteins, 100% of New Zealand white rabbits were resistant to attack of McHDV, which indicates efficient immune-protective efficacy of chosen epitope recombinant protein. The study laid a foundation for the development of the new subunit vaccines of McVHD.

Detection of bluetongue virus in Brazilian cervids in São Paulo state / Detecção de vírus da língua azul em cervídeos brasileiros no estado de São Paulo

Viral hemorrhagic diseases in cervids occur worldwide and include epizootic hemorrhagic disease (EHD), bluetongue (BT), and adenoviral hemorrhagic disease (AHD). Since gross lesions in all three hemorrhagic diseases are identical (hemorrhagic enteropathy, pulmonary edema, systemic petechial and suffusion hemorrhages), it is necessary to use accurate techniques for a definitive etiologic diagnosis. Archival material (paraffin blocks) at the Department of Veterinary Pathology of FCAV - Unesp was reviewed for lesions of hemorrhagic disease and 42 captive and free-living Brazilian deer were selected to include in this study. Paraffin-embedded tissues were evaluated using immunohistochemistry and tested negative for adenovirus. Using real time RT-PCR, EHD virus was not detected in paraffin-embedded tissues in any of the cases evaluated. The same technique was used for detection of BT virus and seven positive animals (16,66%) were confirmed after agarose 4% gel electrophoresis and gene sequencing. The main macroscopic changes observed in the positive animals were hemorrhagic intestinal contents, reddish mucous membrane of the gastrointestinal tract, ulcers on tongue and petechiae in various organs. Microscopic changes observed were lymphocytic inflammatory infiltrate in liver, kidney and lungs, hemorrhage, and congestion in various organs. All positive cases were from captive animals, three females (two young and one adult), and four young males. This study demonstrates that the bluetongue virus is involved in hemorrhagic disease outbreaks of deer in Brazil.(AU)

Doenças hemorrágicas virais em cervídeos ocorrem no mundo todo e incluem a doença epizoótica hemorrágica (DEH), língua azul (LA), e doença hemorrágica por adenovírus (DHA). Uma vez que as lesões nas três doenças hemorrágicas são idênticas (enteropatia hemorrágica, edema pulmonar, petéquias sistêmicas e sufusões hemorrágicas), é necessário utilizar técnicas precisas para um diagnóstico etiológico definitivo. Material de arquivo (blocos de parafina) do Departamento de Patologia Veterinária da FCAV - Unesp foi revisado para lesões de doenças hemorrágicas e 42 cervídeos brasileiros de cativeiro e de vida livre foram selecionados e incluídos neste estudo. Tecidos embebidos em parafina foram avaliados usando imunohistoquímica e foram negativos para adenovírus. Usando o RT-PCR em tempo real, o vírus da DEH não foi detectado nos tecidos de nenhum dos casos avaliados. A mesma técnica foi utilizada para detecção do vírus da LA e sete animais positivos (16,66%) foram confirmados após eletroforese em gel de agarose a 4% e sequenciamento genético. As principais alterações macroscópicas observadas nos animais positivos foram conteúdo intestinal hemorrágico, mucosa do trato gastrointestinal avermelhada, úlceras na língua e petéquias em vários órgãos. As alterações microscópicas observadas foram infiltrado inflamatório linfocítico em fígado, rins e pulmões, e hemorragia e congestão em vários órgãos. Todos os casos positivos foram de animais de cativeiro, três fêmeas (dois jovens e um adulto), e quatro jovens do sexo masculino. Este estudo demonstra que o vírus da lingual azul está envolvido nos surtos de doença hemorrágica em veados no Brasil.(AU)

Evaluation of the proficiency of laboratories for detection of rabbit hemorrhagic disease virus antibody / 中国实验动物学报

Objective Through the detection of rabbit hemorrhagic disease virus( RHDV) antibody, to investigate the capacity of experimental animal quality control laboratories, so as to improve their detection proficiency.Methods According to the program approved by CNAS, the screened samples were numbered randomly and tested for their stability and homogeneity.The random samples were issued to the participant laboratories with the Standard Operation Procedure ( SOP) .The participant laboratories must submit the test reports and original records in time.The feedback results were judged by the rate of concordance with the anticipated results.Results Twenty laboratories from 14 provinces were en-rolled in the evaluation, and all of them submitted detection results on time.ELISA methods were used in 14 laboratories, and hemagglutination inhibition ( HAI) assay was used in 6 laboratories.The results of 17 laboratories were marked as pass or excellent, with a rate of pass of 85%.Conclusions The ability for detection of RHDV antibody in animal test labora-tories in China is high.The implementation of capacity testing can reflect the level of quality control laboratories.

Sequential hepatic ultrastructural changes and apoptosis in rabbits experimentally infected with Korean strain of rabbit hemorrhagic disease virus (RHDVa) / 대한수의학회지

In this study, to understand the pathogenesis of new rabbit hemorrhagic disease virus (RHDVa) serotype, we carried out to administrate RHDVa to rabbits, and to examine sequential electron microscopic changes and relationship between pathogenesis and apoptosis. TUNEL-positive cells began to be observed from 24 hours after inoculation (HAI) and the number of positive cells was slightly increased with the course of time. Whereas marked increase of positive cells was seen in the liver from the rabbits died acutely. Typical viral particles with cup-like projections and a diameter of 30~40 nm were detected in homogenized liver samples and tissues at 36 and 48, and 48 HAI, respectively. Ultrastructurally, glycogen deposition was observed from the first stage of hepatocellular degeneration by RHDVa infection and then, swelling and disruption of cristae of mitochondria by viral particles, swelling of smooth endoplasmic reticulum, vacuoles and vesicles were detected. Condensation, margination and fragmentation of chromatin were observed in degenerative hepatocytes at 36 and 48 HAI, indicating apoptotic bodies. These data offer that hepatocytic apoptosis by RHDV infection could be closely related with mitochondrial impairment in the hepatocytes.

Codon optimization of the rabbit hemorrhagic disease virus (RHDV) capsid gene leads to increased gene expression in Spodoptera frugiperda 9 (Sf9) cells

Rabbit hemorrhagic disease (RHD) is contagious and highly lethal. Commercial vaccines against RHD are produced from the livers of experimentally infected rabbits. Although several groups have reported that recombinant subunit vaccines against rabbit hemorrhagic disease virus (RHDV) are promising, application of the vaccines has been restricted due to high production costs or low yield. In the present study, we performed codon optimization of the capsid gene to increase the number of preference codons and eliminate rare codons in Spodoptera frugiperda 9 (Sf9) cells. The capsid gene was then subcloned into the pFastBac plasmid, and the recombinant baculoviruses were identified with a plaque assay. As expected, expression of the optimized capsid protein was markedly increased in the Sf9 cells, and the recombinant capsid proteins self-assembled into virus-like particles (VLPs) that were released into the cell supernatant. Rabbits inoculated with the supernatant and the purified VLPs were protected against RHDV challenge. A rapid, specific antibody response against RHDV was detected by an ELISA in all of the experimental groups. In conclusion, this strategy of producing a recombinant subunit vaccine antigen can be used to develop a low-cost, insect cell-derived recombinant subunit vaccine against RHDV.

Sequential pathologic changes and viral distribution in rabbits experimentally infected with new Korean strain of rabbit hemorrhagic disease virus (RHDVa) / 대한수의학회지

Rabbit hemorrhagic disease is a highly acute and fatal viral disease caused by rabbit hemorrhagic disease virus (RHDV). Since first outbreak in Korea 1987, RHDV has been continually affected in the country, but the pattern of outbreak seem to be changed. In this study, to understand the pathogenesis of the new RHDVa serotype, we therefore carried out to inoculate RHDVa to rabbits, and to examine the sequential histopathologic changes and viral distribution. Macroscopically, various sized dark red or white spots or appearance were observed in the liver, lung, kidney uterus and ureter. In euhanized rabbits, significant pathologic findings such as infiltration of heterophils and mononuclear cells were observed at 24 hours after inoculation (HAI), and these were sequentially extended periportal to centrilobular area. However, in dead rabbits, severe hepatic degeneration and/or necrosis with relatively weak inflammatory responses were observed. RHDV antigens began to detect in liver, spleen, and lung from 12 HAI by PCR. Immunohistochemically, RHDV positive cells were seen in only liver from 24 HAI, and the degree of immunogen reactivity was stronger in dead rabbits than in euthanized ones. In conclusion, RHDVa caused the subacute or chronic infection accompanying low mortality and moderate to severe inflammatory reaction in rabbits, suggesting the possibility that RHD could become endemic.

Clinical analysis of 137 pediatric cases with coagulation disorders / 中国小儿急救医学

Objective To analyze the pathogenesis,therapy and outcome of pediatric cases with coagulation disorders (CD).Methods All these 137 patients were diagnosed as CD with the methods of hemoglutination five items and/or disseminated intravascular coagulation indexes.Then activity of specific coagulation factors,morphology of bone marrow,hepatorenal function and some other relative tests were performed to find out the cause of CD or the primary disease.Results Forty-three cases were diagnosed as genetic CD with 29 as hemophilia A,4 as hemophilia B and 10 as Von Willebrand disease;while the other 94 patients as acquired CD with 15 as vitamin K-dependence coagulation factor deficiency,22 as hepatic dysfunction,30 as disseminated intravascular coagulation and 1 as thrombotic thrombocytopenic purpura.Genetic CD was treated with replacement therapy to reduce the complication.There was 1 case in this group died of intracranial hemorrhage.Acquired CD was treated with short-term,specific and necessary replacement therapy on the basis of reasonable treatment of primary diseases.Eleven cases died finally in this cohort with 7 cases as liver failure and the other 4 cases as terminal leukemia or lymphoma.Conclusion Pediatric patients with CD were caused by genetic or acquired diseases.In clinic the reason of CD was mainly acquired.The treatment of genetic CD is the replacement of specific coagulation factor for life-long term.The outcome dependes on the lack of degree.While the therapy for acquired CD aims at the primary disease.The principle of blood transfusion is short-term and the outcome dependes on the therapic effects of primary diseases.

Granulomatous myelitis associated with hemorrhagic syndrome due to consumption of Vicia villosa by cattle / Mielite granulomatosa associada à síndrome hemorrágica devido ao consumo de Vicia villosa em um bovino

An unusual case of spontaneous Vicia villosa poisoning affected a 6-year-old Holstein cow. Although the most striking findings included a generalized hemorrhagic condition associated with granulomatous myelitis, histological lesions typically seen with the vetch-associated systemic granulomatous syndrome were also present. Prominent gross findings were bloody nasal and oral discharges, disseminated hemorrhages, and bloody feces. Generalized hemorrhages associated with infiltration of numerous organs by lymphocytes, plasma cells, macrophages, multinucleated giants cells, and eosinophils were the main microscopic findings. Anti-CD68 immunostaining confirmed the presence of moderate histiocytic infiltrate and multinucleated giant cells in the bone marrow. These changes in the bone marrow probably caused the generalized hemorrhagic changes described here.

Um bovino Holandês de seis anos naturalmente intoxicado pela Vicia villosa apresentou lesões não comumente encontradas nessa intoxicação. Embora a lesão mais evidente fosse hemorragia generalizada associada à mielite granulomatosa, as lesões granulomatosas típicas da intoxicação pela ervilhaca também estavam presentes histologicamente. Os principais achados macroscópicos foram hemorragias nasal e oral, hemorragias generalizadas e fezes com sangue. Lesões hemorrágicas generalizadas associadas com infiltrado inflamatório de linfócitos, plasmócitos, macrófagos, células gigantes multinucleadas e eosinófilos foram os principais achados microscópicos encontrados. O teste imuno-histoquímico anti-CD68 confirmou a presença de infiltrado moderado de macrófagos e de células gigantes multinucleadas na medula óssea. As alterações na medula óssea provavelmente constituem a causa da hemorragia generalizada descrita nesse caso.

Pathological anatomy study of a hemorrahagic disease in four (4) pigs

Hay muchas enfermedades con hemorragias difusas que ocurren en cerdos alrededor del mundo. Estas pueden ser de origen viral como el Cólera Porcino (C P) conocido como Fiebre Porcina Clásica ( F P C) o Peste Porcina Clásica (P P C), producido por un Pestivirus (ARN) del grupo Togavirus, de la familia Flaviviridae; o también la Peste Porcina Africana (P P A) producida por un Iridovirus y la cual de acuerdo a la literatura no ha sido diagnosticada en Venezuela. Entre las de origen bacteriano están la Salmonelosis y Erisipelosis porcina, siendo ésta última poco frecuente en Venezuela. Los cambios hemorrágicos observados en los cerdos bajo estudio son muy similares a aquellos que ocurren en la F P C caracterizada ésta, por afectar varios órganos vitales e induciendo una alta tasa de mortalidad. La F C P cursa en forma aguda con hemorragias generalizadas, alta morbilidad y mortalidad o en forma crónica con signos clínicos y lesiones mínimas o ausentes. El agente etiológico de F P C tiene un efecto letal en animales infectados de forma natural y en animales postvacunados induce lesiones letales, que aunque más suaves producen una alta mortalidad, debido a factores de la vacuna o al estatus inmunológico del cerdo...

Many diseases occur in pigs causing widespread hemorrhages around the world. Among them are viral or bacterial such as, Classical Swine Fever (C S F) which is produced by a Pestivirus (RNA), Togavirus group of the Flaviviridae family, besides African Swine Fever (ASF) produced by an Iridovirus and which has not been diagnosed in Venezuela, according to the literature. Among bacterial diseases are Salmonellosis and Erysipellosis, being the latter uncommon in Venezuela. In this paper, the morphological aspects of a hemorrhagic disease in four pigs are described. The reported lesions are not too dissimilar to those occurring in cases of C S F, which is characterized by affecting many vital organs, producing death in many pigs. C S F often follows an acute course characterized by generalized hemorrhages, with high morbidity and mortality, but it may be chronic, in that case clinical signs and lesions are often minimal or absent. The disease in naturally infected pigs produces lethal lesions causing death in many unvaccinated animals as well as in vaccinated pigs. The gross and microscopic changes of a hemorrhagic systemic disease in four (4) cases under study are reported from two naturally infected unvaccinated pigs and two postvaccinated animals against C S F. Pathologic changes are correlated with clinical signs and the most relevant morphological gross changes and histopathology are reported from naturally infected unvaccinated and postvaccinated pigs. The hemorrhagic lesions reported in vital organs highly resemble and are very similar to those morphologic changes occurring in C S F, although, other hemorrhagic viral or bacterial diseases induced a more severe and different pattern in lesions though being hemorrhagic. Despite lab tests (Elisa) were not determined in this work, they must be ruled out to show the etiological agent of C S F in spite of the typical and very similar morphological changes reported in this paper.

Investigação de surto de doença febril hemorrágica nos municípios de Japeri e Queimados, Rio de Janeiro, 2000 / Outbreak investigation of hemorragic febrile disease in Japeri and Queimados, Rio de Janeiro, 2000

Entre junho e setembro de 2000, 9 pessoas de uma área pobre da zona norte do Rio de Janeiro, Brasil, desenvolveram uma doença febril hemorrágica e morreram. Foi conduzida uma busca ativa para determinar a extensão do surto e um estudo de caso-controle para identificar a etiologia e os fatores de risco para aquisição da doença. Um paciente-caso foi definido como residente da área afetada apresentando febre, cefaléia e um ou mais dos seguintes sintomas - mialgia, artralgia, diarréia, manifestações hemorrágicas ou icterícia - durante o período de junho a setembro de 2000, tendo procurado serviço médico. Quarenta e três pessoas enquadraram-se na definição de caso e foram comparados com 86 controles, pareados por área de residência e faixa etária. Vinte e três (53,3%) eram do sexo masculino e a idade mediana foi de 19 anos. Na análise univariada, contato com esgoto (mOR=2,5; p=0,02); contato com ratos (mOR=3,0; p=0,006), contato com rio (mOR=3,3; p=.009), e mortandade de animais no peridomicílio (mOR=2,7; p=0,04), foram significantemente associados ao desenvolvimento de doença. Na análise multivariada, contato com ratos (OR=5,1; p=0,03) e contato com rio (OR=4,9; 95%; p=0,04) permaneceram como fatores de risco independentemente associados ao desenvolvimento de doença. A imunohistoquímica de uma amostra de tecido de um caso fatal foi positiva para leptospirose. Exames adicionais para outros potenciais agentes estão pendentes. Com base nas evidências epidemiológicas fornecidas pela investigação, recomendamos a implementação de desratização na área afetada e educação em saúde aos residentes, para que evitassem contato com o rio local. Após a implementação das medidas de controle nenhum caso novo foi notificado. Esta investigação ressalta a importância de se instituir recomendações de saúde pública com base em resultados de investigações epidemiológicas quando exames laboratoriais não estão prontamente disponíveis.

In June-September 2000, nine persons from a poor area north of Rio de Janeiro, Brazil, developed a febrile hemorrhagic illness and died. We conducted case finding to determine the outbreak size, and a case-control study to identify disease etiology and risk factors. A case-patient was defined as a resident of the affected area who sought medical care during June-September 6, 2000, presenting with fever, headache, and one or more of the following symptoms: myalgia, arthralgia, diarrhea, hemorrhagic manifestations, or jaundice. Forty-three persons met the case definition and were compared with eighty-six controls matched by area of residence and age group. Twenty-three (53.3%) were males with a median age of 19 years. In univariate analysis, contact with sewage (matched Odds Ratio [mOR]=2.5; p=0.02), contact with rats (mOR=3.0; p=0.006), contact with local creek (mOR=3.3; p=0.009), and mortality of household animals (mOR=2.7; p=0.04), were associated with development of the disease. In multivariate analysis, contact with rats (mOR=4.7; p= 0.03), and contact with local creek (mOR=4.8; 95%; p= 0.04) remained as independent risk factors. Immunohistochemical evaluation of tissue samples from one fatal case was positive for leptospirosis. Further laboratory testing for potential etiologies is pending. Based on the epidemiologic evidence provided by our investigation we recommended implementation of rodent extermination in the affected area, and education of residents to avoid contact with the local creek. Following implementation of these control measures, further cases were not reported. This investigation highlights the value of implementing public health recommendations based on results from epidemiologic investigations when laboratory testing is not immediately available.

Intracranial Hemorrhage in Late Hemorrhagic Disease of the Newborn: Brain Computed Tomographic Findings / 대한소아신경학회지

PURPOSE: This study is to evaluate the Brain CT manifestations of late hemorrhagic disease of the newborn. METHODS: We evaluated 13 cases with late hemorrhagic disease of the newborn, for whom brain CT scans were performed. Clinical indications for brain CT scan were mental changes(n=5), vomiting(n=2), irritability(n=2), seizure(n=3), and lethargy(n=1). We analyzed the result of the brain CT findings with attention to anatomic locations of hemorrhage and characteristics of hemorrhagic manifestations. RESULTS: Only one patient in 13 patients was normal on brain CT scan, and 12 patients showed hemorrhagic lesions. Among 12 patients, 5 cases had single hemorrhagic lesion(3 subdural hemorrhages and 2 intracerebral hemorrhages, and 7 cases had multiple lesions, in which subdural hemorrhage and subarachnoid hemorrhage were common respectively. The other common brain CT findings except hemorrhagic lesion were the mass effects with ventricle compression(n=11), midline shifting(n=6), ventricular dilatation(n=7), and fluid-fluid levels in hemorrhagic lesion(n=9). CONCLUSION: The common intracranial hemorrhages of late hemorrhagic disease of the newborn were subdural and subarachnoid hemorrhages. The associated CT findings were mass effect and fluid-fluid levels. Intracranial hemorrhage in late hemorrhagic disease of newborn could massively occurr. Thus brain CT scan should be done if any symptom for neurologic abnormality was present.

Prophylactic Effect of Intramuscular, Oral, and Maternal Administration of Vitamin K on Hemorrhagic Disease of the Newborn

PURPOSE: Vitamin K deficiency is associated with hemorrhagic disease of the newborn. Late hemorrhagic disease is often intracranial and may be fatal. Many countries recommend vitamin K prophylaxis after birth to prevent this hazard of vitamin K deficiency. Nevertheless, there are still controversies concerning the best way of providing effective prophylaxis. A recent article by Golding and colleagues has questioned the safety of the routine use of intramuscular vitamin K for the newborn. These authors reported a significantly increased rate of childhood cancer in infants who received intramuscular prophylaxis. So we compared the prophylactic effect of intramuscular, oral, and maternal administration of vitamin K on hemorrhagic disease of the newborn. METHODS: A total of 60 newborns, delivered spontaneously vaginally, in the Masan Fatima hospital from March to June, 1996, were enrolled. Neonated with intrapartum anoxia, liver disease or hereditary coagulation factor deficiencies, who received antibiotics were excluded. Mothers receiving any medication known to interferes with vitamin K metabolism(such as antiepileptics, antibiotics and anticonvulsions) were excluded. The newborns were randomly allocated to one of the four groups. A group was not supplied. B group received 1mg of vitamin K1 intramusculary, C group received 2mg of vitamin K1 orally. D group was given 20mg of vitamin K1 orally to their mothers at least 2days(range 2 to 7) before birth. Blood samples were collected from 48hrs to 72hrs after birth. PIVKA-II level was measured by enzyme-linked immunosorbent assay (EITEST-MONOP, Eisai Ltd), using a monospecific monoclonal antibody against PIVKA-II. The results obtained are expressed in arbitrary unit (AU) : 1AU corresponds to 1micro gram of purified prothrombin. (healthy adults have less than 0.13AU/ml). PT, PTT were measured simultaneously. RESULTS: 1) PIVKA-II was detected in 4 of 15 infants in group A, who were not supplied. None was detected in other groups. So PIVKA-II detection rate was significantly decreased in other groups compared with group A(p<0.05). 2) PT(sec) values were 12.74+/-0.91, 12.58+/-0.89, 12.36+/-1.04, 12.16+/-0.90 respectively, and there was no significant difference between groups. 3) PTT(sec) values were 52.41+/-13.26, 38.39+/-10.04, 42.67+/-7.01, 39.77+/-10.48 respectively and there was significant shortening in other groups compared with group A (p<0.05). CONCLUSION: Not only intramuscular administration but oral and maternal administration of vitamin K have prophylactic effect on hemorrhagic disease of the newborn. Prophylactic effect on the late hemorragic disease of the newborn requires further extensive study and evaluation.

A Case of Combined Congenital Deficiency of Factor V and Factor VIII

Hemorrhagic disease of newborn by combined blood clotting factor deficiencies is very rare. Combined deficiency of factor V and factor VIII is the most common form among these cases, and inherited by autosomal recessive trait. Clinical findings are easy bruising, post-traumatic bleeding and bleeding after tooth extractions, and the main laboratory findings are prolonged partial thromboplastin time (PTT) and prothrombin time (PT) that are different from hemophilia A only with prolonged PTT. We experienced a case of combined congenital deficiency of factor V and factor VIII in a 1-day-old boy. We report the case with brief review of literature.

Vitamin K-Deficient Hemorrhagic Disease in Infants with Acute Subdural Hematoma

We had experienced 2 cases of acute subdural hematoma due to vitaimin K(vit. K) deficiency. They were 44-and 42-day-old. After the adminstration of vit.K1, prolonged prothrombin time(PT) and activated partial thromboplastin time(APTT) were corrected, so the surgical treatment ws performed. We supposed the predisposing factors were breast feeding and unknown hepatic pathology. No complication remained to one infant but the other multiple cerebral infarctions. The etiology, pathogenesis, diagnosis, prevention are discussed. We insist on neonatal administration of vit. K1 for the prevention of bleeding tendency. If intracranial hemorrhage develops, we must keep in mind that the early neurosurgical intervention may be needed.

Late Hemorrhagic Disease of Infancy due to Vitamin K Dependent Factor Deficiency

A clinical study was done on 9 children with 10 episodes, who were admitted to our pediatric ward under the diagnosis of late hemorrhagic disease of infancy due to vitamin K dependent factor deficiency. Study period was past 2 years, from September 1979 to May 1981. The following results were obtained. This disorder occurred most frequently during early infancy, with male predominence, male female ratio being 4:1. And breast feeding was noted in 6, with history of previous medication for URI in 7. GIT was the most common site of bleeding occurring in 9 cases, followed by CNS in 3, skin in 1 and nose in 1. Anemia, probably due to blood loss, was seen in 8 cases, and dehydration, oral thrush, fever, irritability, mental change and convulsion were seen in small number of cases. Initial laboratory findings showed prolonged PT and PTT with normal platelet count in all cases. After vitamin K therapy, PT and PTT became normal in majority of cases. Prompt response to vitamin K therapy within 24 hours was noted in 6, prolonged response over 3 days in 2, and one patient showed recurrence at 2(1/2) months after complete recovery from first episode.

Five Cases of Hemorrhagic Disease of Infancy

A hemorrhagic disease due to acquired prothrombin complex deficiency is presented in five infants from 3 weeks to 2 months of age. There are 2 cases of intracranial hemorrhage, 1 case of hemothorax, 2 cases of gastrointestinal bleeding, 1 case of epistaxis and 3 cases of petechia, purpura or bleeding from injection site. After the administration of vitamin K, bleeding ceased and prolonged prothrombin time was corrected promptly. Vitamin K dependent coagulation factor deficiency due to vitamin K deficiency is accounted for the pathogenesis of bleeding. Diarrhea, treatment with antimicrobial agents and a diet low in vitamin K are suggested as etiologic factors acting together.

Five Cases of Hemorrhagic Disease of Infancy

A hemorrhagic disease due to acquired prothrombin complex deficiency is presented in five infants from 3 weeks to 2 months of age. There are 2 cases of intracranial hemorrhage, 1 case of hemothorax, 2 cases of gastrointestinal bleeding, 1 case of epistaxis and 3 cases of petechia, purpura or bleeding from injection site. After the administration of vitamin K, bleeding ceased and prolonged prothrombin time was corrected promptly. Vitamin K dependent coagulation factor deficiency due to vitamin K deficiency is accounted for the pathogenesis of bleeding. Diarrhea, treatment with antimicrobial agents and a diet low in vitamin K are suggested as etiologic factors acting together.