In Vitro Activities of MMV Malaria Box Compounds against the Apicomplexan Parasite Neospora caninum, the Causative Agent of Neosporosis in Animals.

(1) Background: Neospora caninum is a major cause of abortion in cattle and represents a veterinary health problem of great economic significance. In order to identify novel chemotherapeutic agents for the treatment of neosporosis, the Medicines for Malaria Venture (MMV) Malaria Box, a unique collection of anti-malarial compounds, were screened against N. caninum tachyzoites, and the most efficient compounds were characterized in more detail. (2) Methods: A N. caninum beta-galactosidase reporter strain grown in human foreskin fibroblasts was treated with 390 compounds from the MMV Malaria Box. The IC50s of nine compounds were determined, all of which had been previously been shown to be active against another apicomplexan parasite, Theileria annulata. The effects of three of these compounds on the ultrastructure of N. caninum tachyzoites were further investigated by transmission electron microscopy at different timepoints after initiation of drug treatment. (3) Results: Five MMV Malaria Box compounds exhibited promising IC50s below 0.2 µM. The compound with the lowest IC50, namely 25 nM, was MMV665941. This compound and two others, MMV665807 and MMV009085, specifically induced distinct alterations in the tachyzoites. More specifically, aberrant structural changes were first observed in the parasite mitochondrion, and subsequently progressed to other cytoplasmic compartments of the tachyzoites. The pharmacokinetic (PK) data obtained in mice suggest that treatment with MMV665941 could be potentially useful for further in vivo studies. (4) Conclusions: We have identified five novel compounds with promising activities against N. caninum, the effects of three of these compounds were studies by transmission electron microscopy (TEM). Their modes of action are unknown and require further investigation.

Socio-economic burden of parasitic infections in yaks from 1984 to 2017 on Qinghai Tibetan Plateau of China-A review.

Yak is an important animal for the Tibetans at Qinghai-Tibetan Plateau of China. The burden of parasitic diseases has been a major threat to the health of yaks at this region presenting a considerable socio-economic losses and impact to yak production and local nomads. Keeping in view, we collected the published papers from 1984 to 2017 on major parasitic infections in yaks by electronic literature search from five databases including CNKI, Google, PubMed, Science Direct and Web of Science. The prevalence of Eimeria, Babesia, Theileria, Hypodermosis, Cystic echinococcosis, Alveolar echinococcosis, Toxoplasma gondii, Neospora caninum, Cryptosporidium, Giardia duodenalis, Enterocytozoon bieneusi, Toxocara vitulorum, and Fascioliasis infection in yaks was found to be 48.02%, 13.06%, 36.11%, 59.85%, 16.93%, 0.99%, 20.50%, 5.14%, 10.00%, 3.68%, 4.07%, 22.23% and 28.7% respectively. Data presented are contemplated to enhance our current understanding on the major parasitic diseases of yaks at Qinghai Tibetan plateau, China. The main aim of this effort is to ameliorate the effects of the parasitic burden in this specie; so that, the attempts are made to minimize the incidence of these infections in future to raise the socio-economic levels of local community.

Placental lesions associated with abortion and stillbirth in goats naturally infected by Neospora caninum / Lesões placentárias associadas a abortos e natimortos em cabras naturalmente infectadas pelo Neospora caninum

Neospora caninum has been described as a parasite that sporadically causes reproductive problems in goats. Several aspects of the pathogenesis of neosporosis in naturally infected goats remain to be established. The aims of the present study were to characterize the placental lesions in goats naturally infected by N. caninum and to evaluate several diagnostic techniques for effective detection of this protozoan in the goat placenta. Some placentas in this study originated from abortion and stillbirth in which there were severe lesions. The lesions were characterized mainly by necrosis involving the mesenchyme of the chorionic villi and trophoblast cells often alongside mononuclear inflammation and in some cases with neutrophilic infiltration. N. caninum DNA was detected in these placentas, but parasite structures were not visualized through immunohistochemistry (IHC). However, five of 11 placentas from N. caninum-infected goats that gave birth to healthy kids had histological lesions characterized by mononuclear inflammation. Of these 11 placentas, N. caninum DNA was detected in seven, and N. caninum tachyzoites were detected in only one of these seven placentas using IHC. The present study demonstrates that severe lesions in the placenta are associated with abortion and stillbirth in caprine neosporosis and the placental alterations are likely involved in abortion pathogenesis. Moreover, the results highlight the importance of using more than one diagnostic technique for the detection of the protozoan in placentas because N. caninum cannot be reliably detected by histological and immunohistochemical tests.(AU)

Neospora caninum é descrito como um parasito que causa problemas reprodutivos esporádicos em cabras. Muitos aspectos da patogênese da neosporose em cabras naturalmente infectadas ainda precisam ser estabelecidos. Os objetivos deste trabalho foram caracterizar as lesões placentárias em cabras naturalmente infectadas por N. caninum e avaliar as técnicas diagnósticas para a detecção efetiva do protozoário na placenta. Algumas placentas deste estudo são originárias de abortos e natimortos, nas quais havia lesões graves. As lesões foram classificadas principalmente por necrose envolvendo o mesênquima das vilosidades coriônicas e células trofoblásticas, geralmente associadas a infiltrado inflamatório mononuclear e em alguns casos infiltrado neutrofílico. O DNA do N. caninum foi detectado nestas placentas, porém estruturas parasitárias não foram visualizadas na imuno-histoquímica (IHQ). Entretanto, cinco das onze placentas de cabras infectadas, que deram à luz a conceptos saudáveis, apresentaram lesões histológicas caracterizadas por infiltrado inflamatório mononuclear. Destas 11 placentas, foi detectado DNA de N. caninum em sete e taquizoítos foram encontrados em apenas uma por meio de imuno-histoquímica. O presente estudo demonstra que abortos e natimortos na espécie caprina, causados pelo N. caninum estão associados a lesões acentuadas nas placentas, sendo que as mesmas estão envolvidas na patogênese do aborto. Os resultados também ressaltam a importância do uso de mais de uma técnica diagnóstica para a detecção do protozoário em placentas, pois o N. caninum não pode ser confiavelmente detectado somente pelos exames de histopatologia e imuno-histoquímica.(AU)

Towards a preventive strategy for neosporosis: challenges and future perspectives for vaccine development against infection with Neospora caninum.

Neosporosis is caused by the intracellular protozoan parasite Neospora caninum. This major disease-causing pathogen is responsible for inducing abortion in cattle, and these adverse events occur sporadically all over the world, including Japan. Currently, there are no vaccines on the market against infection with N. caninum. Because live and attenuated vaccines against N. caninum have had safety and effectiveness issues, development of a next-generation vaccine is urgently required. To develop a vaccine against neosporosis, my laboratory has been focused on the following: 1) understanding the host immune responses against Neospora infection, 2) identifying vaccine antigens and 3) developing an effective antigen-delivery system. The research strategy taken in my laboratory will have strong potential to progress current understanding of the pathogenesis of N. caninum infection and promote development of a novel subunit vaccine based on the specific vaccine antigen with an antigen-delivery system for controlling neosporosis.

Desempenho reprodutivo entre receptoras bovinas não contaminadas e naturalmente infectadas por Neospora caninum, após a transferência de embriões / Reproductive performance in bovine receipts non infected and naturally infected with Neospora caninum after embryo transfer

O uso do conhecimento de biotécnicas para a melhoria da produção de bens e serviços tem avançado significativamente. O controle de doenças infecciosas continua a representar o maior obstáculo para a saúde dos animais. A perda embrionária pode ser responsável pelo aumento de prejuízos econômicos isolados para os produtores de bovinos. A infecção por Neospora caninum tem emergido como uma importante doença reprodutiva e, em vários países, tem sido diagnosticada como principal causa de aborto. O objetivo desse trabalho foi comparar o desempenho reprodutivo em receptoras bovinas não contaminadas e naturalmente infectadas por Neospora caninum após a transferência de embriões (TE). A escolha dos animais (n=395) para o experimento foi realizada através de avaliação ginecológica, via palpação transretal. As vacinações seguiram protocolo sanitário recomendado para a TE. O manejo nutricional foi baseado no National Research Council (NRC, 1999). A TE de embriões descongelados foi feita após a sincronização do ciclo estral das receptoras. Para a avaliação do desempenho reprodutivo se utilizou de ultrassonografia transretal. A identificação das receptoras positivas para Neospora caninum realizou-se através do teste de ELISA e, para as amostras reagentes, confirmação através de imunofluorescência indireta (IFI). Os dados foram analisados utilizando Past® e foram considerados estatisticamente significantes (P<0,01). Os resultados mostram uma taxa de prenhes (n=191) de 48,35%, seguida de uma taxa de abortos aos 60 dias de gestação (n=14) de 3,5% e abortos ao sexto mês de gestação (n=12) de 3,04%. Esses resultados tornam-se relevantes quando se leva em consideração que, receptoras infectadas pelo protozoário não apresentaram uma taxa elevada de perdas gestacionais. A insignificância estatística dos dados obtidos nesse estudo confere a segurança em sugerir que não há necessidade em implantar o diagnóstico ou controle prévios para Neosporose em rebanhos aptos a TE.

The use of knowledge in biotechnologies for improving the production of goods and services has advanced significantly. The control of infectious diseases continues to represent the most obstacle to the health of animals. The embryonic loss may be responsible for the increase in economic losses for producers isolated from cattle. Infection with Neospora caninum has emerged as an important reproductive disease and, in several countries, has been diagnosed as the main cause of abortion. The aim of this study was to evaluate the reproductive performance in not contaminated and naturally infected with Neospora caninum bovine receipt after embryos transfer (ET). The choice of animals (n=395) for the experiment was performed by gynecological evaluation, via transrectal palpation. Vaccinations recommended for health protocol followed the ET. Nutritional management was based on the National Research Council (NRC, 1999). The ET of thawed embryos was made after the synchronization of the estrous cycle of the receipts. To evaluate the reproductive performance of transrectal ultrasonography was used. The identification of the positive receipts for Neospora caninum was performed by ELISA and reagents for samples, confirmation by indirect immunofluorescence (IIF). Data were analyzed using Past® and were considered statistically significant (P<0.01). The results show a rate of pregnant (n=191) of 48.35%, followed by an abortion rate at 60 days of gestation (n=14) and 3.5% of abortions in the sixth month of gestation (n=12) of 3.04%. These results become relevant when taking into consideration that infected receipts by the protozoan not showed a high rate of pregnancy loss. The statistical insignificance of the data obtained in this study provides the security suggesting that there is no need to deploy the diagnosis or prior control to neosporosis in receipts able to ET.

Toxoplasmosis and neosporosis among beef cattle slaughtered for food in Western Thailand.

Beef is a main type of meat consumed by Thais. The prevalences of anti-Toxoplasma gondii and anti-Neospora caninum antibodies were investigated among beef cattle slaughtered for food in western Thailand. A total of 389 blood samples obtained from beef cattle from 24 herds were collected at 3 slaughterhouses in 3 western provinces of Thailand: Kanchanaburi, Ratchaburi and Nakhon Pathom. An indirect immunofluorescent antibody test (IFAT) was performed using cut-off values of 1:128 for T. gondii and 1:200 for N. caninum. The antibodies to T. gondii were found in 100 samples (25.7%) and antibodies to N. caninum were found in 23 samples (5.9%) a significant difference (p < 0.001) in prevalences, indicating the cattle tested had a greater exposure to T. gondii than N. caninum, and they should be regarded as a potential source of T. gondii infection to humans. The low prevalence of neosporosis in this study is still a risk for morbidity among cattle, including abortions. This is the first study in Thailand finding both T. gondii and N. caninum antibodies among beef cattle.

Occurrence of anti-Neospora caninum antibodiesin dogs in rural areas in Minas Gerais, Brazil / Ocorrência de anticorpos anti-Neospora caninum em cães de áreas rurais em Minas Gerais, Brasil

The aim of this study was to determine the frequency of anti-Neospora caninum antibodies and risk factors associated with seropositivity in 240 dogs from rural areas of the Lavras, Belo Horizonte and Nanuque regions, state of Minas Gerais, Brazil. The indirect fluorescence antibody test (IFAT) with a cutoff of 50 was used to assess the frequency of seropositive dogs. The risk factor analysis was performed using the Chi-square test (χ2) and multiple logistic regression. The frequency of seropositivity for N. caninum was 15% of the dogs (36/240). Presence of dogs without defined breed (p = 0.018; OR = 5.9) and presence of cattle on the farm (p = 0.053, OR = 4.3) were associated with N. caninum seropositivity. Dogs in the Nanuque region had higher seropositivity for N. caninum (29.9%) than did those in the Lavras (6.2%) and Belo Horizonte (2.2%) regions (p < 0.05). The presence of seropositive dogs in rural areas of Minas Gerais demonstrates the potential risk of horizontal transmission of N. caninum to cattle, especially from dogs without defined breed, which were four times more likely to be seropositive than purebred dogs were.

O objetivo deste estudo foi determinar a frequência de anticorpos anti-Neospora caninum e fatores de risco associados à soropositividade, em 240 cães de áreas rurais, nas regiões de Lavras, Belo Horizonte e Nanuque, Minas Gerais, Brasil. Foi utilizada a reação de imunofluorescência indireta (RIFI), com um ponto de corte igual a 50, para avaliar a frequência de cães soropositivos. A análise dos fatores de risco foi realizada pelo teste de Qui-quadrado (χ2) e regressão logística múltipla. A frequência de cães soropositivos para N. caninum foi de 15% (36/240). A presença de cães sem raça definida (SRD) (p = 0,018; OR = 5,9) e bovinos na propriedade (p = 0,053; OR = 4,3) foi associada à soropositividade pelo N. caninum. A frequência de cães soropositivos foi mais elevada na região de Nanuque (29,9%), quando comparados aos das regiões de Lavras (6,2%) e Belo Horizonte (2,2%) (p < 0,05). A presença de cães soropositivos, em áreas rurais de Minas Gerais, demonstra o risco potencial da transmissão horizontal de N. caninum para os bovinos, especialmente os cães SRD, que apresentaram quatro vezes mais a chance de serem soropositivos em relação aos cães com raça definida.

Quantification of vertical transmission of Neospora caninum in dairy cows in Minas Gerais, Brazil / Quantificação da transmissão vertical de Neospora caninum em vacas leiteiras de Minas Gerais, Brasil

The objective of this cross-sectional study was to estimate the rate of vertical transmission and to investigate horizontal transmission of Neospora caninum and occurrences of reproductive abnormalities in seropositive dairy cows on two farms in the state of Minas Gerais, Brazil. The frequency of cows seropositive for N.caninum according to the indirect fluorescent antibody test (IFAT) was 39.4% (93/236) for Farm A and 31.4% (32/102) for the Farm B (p > 0.05). The mean vertical transmission rates for N. caninum were 29% and 9% for the herds of Farms A and B, respectively. No negative effects (p > 0.05) from infection by N.caninum were observed regarding milk production and occurrences of reproductive abnormalities in herds A and B.

O objetivo deste estudo transversal foi estimar a taxa de transmissão vertical e investigar a transmissão horizontal de Neospora caninum, e a ocorrência de alterações reprodutivas em vacas leiteiras soropositivas em duas fazendas no Estado de Minas Gerais, Brasil. A frequência de vacas soropositivas a N. caninum pelo teste de imunofluorescência indireta (RIFI) foi de 39,4% (93/236) e 31,4% (32/102) para as fazendas A e B, respectivamente. A taxa média de transmissão vertical de N. caninum, respectivamente, foi de 29% e 9% para os rebanhos A e B. Não foi observado nenhum efeito negativo (p > 0,05) da infecção por N. caninum sobre a produção de leite e a ocorrência de alterações reprodutivas nos rebanhos A e B.

Fatores associados a soroprevalência de neospora caninum e toxoplasma gondii em rebanhos caprinos na região sul de Minas Gerais / Factors associated to seroprevalence of neospora caninum and toxoplasma gondii in caprine herds in southern Minas Gerais state, Brazil

Este estudo estabelece a soroprevalência de Neospora caninum e Toxoplasma gondii em rebanhos caprinos no Sul de Minas Gerais e verifica a sua associação com o desempenho reprodutivo dos rebanhos analisados, uma vez que não há dados sobre neosporose em caprinos neste Estado. Amostras de soro de 401 caprinos provenientes de 11 propriedades foram submetidas à reação de imunofluorescência indireta anti-N. caninum e T. gondii. A prevalência média de animais positivos foi de 10,7% e 21,4%, respectivamente. Houve diferença significativa entre as faixas etárias. Neste quesito os maiores índices de positividade, para ambos os parasitas, foram observados nos caprinos acima de três anos de idade. Estes apresentaram uma probabilidade 2,6 e 4,8 maior, em relação aos animais de até um ano de idade, para N. caninum e T. gondii, respectivamente. No teste T não houve associação positiva entre as médias de prevalência de N. caninum (p=0,553) e T. gondii (p=0,098) com a ocorrência de problema reprodutivo nos rebanhos. Porém, propriedades com histórico de problemas reprodutivos apresentaram uma média de reagentes para N. caninum de 18,64%, se comparadas a 13,97% de criatórios que não relataram essas alterações. Quando os problemas reprodutivos foram separados em categorias a relação entre a ocorrência de aborto e a soroprevalência de N. caninum e de T. gondii foi significativa (p<0,05) por meio dos testes Exato de Fisher e Qui-quadrado. Não houve diferença (p>0,05) entre as médias de prevalência de T. gondii e o acesso de gatos ao rebanho no teste T. Entretanto, a média de prevalência nas propriedades em que os gatos tinham acesso foi de 31,01%, se comparada a 17,34% em criatórios onde não existiam gatos. Foi constatado o acesso de cães em 100% das propriedades. Os resultados obtidos neste estudo demonstram que N. caninum e T. gondii infectam caprinos no Sul de Minas Gerais e que estes parasitas podem estar associados à ocorrência de abortos nessa espécie animal.

This study determines the seroprevalences of Neospora caninum and Toxoplasma gondii in caprine herds in southern Minas Gerais state and investigates their association with the reproductive performance of the herds analyzed since no data concerning neosporosis in goats in this state are available. Serum samples from 401 goats from 11 farms were subjected to indirect immunofluorescence reaction for the detection of N. caninum and T. gondii. The mean prevalence rates of animals that tested positive were 10.7% and 21.4%, respectively. Significant differences between different age groups were observed. The highest frequencies of positive cases for both parasites were observed in goats over three years of age; in this group, the probability of seropositivity for N. caninum and T. gondii was 2.6 and 4.8 times higher, respectively, when compared to animals up to one year of age. No positive correlation was found between the mean prevalence rate of N. caninum (p-0,553) and T. gondii (p=0.098) and the occurrence of reproductive problems in the herds using T-test. However, farms with a history of reproductive problems exhibited an average of 18.64% animals reactive to N. caninum, compared to 13.97% from farms where those problems were not reported. When reproductive problems were separated in category, the relationship between occurrence of abortion and seroprevalence of N. caninum and T. gondii was significant (p<0.05) using Fisher’s exact test and chi-square test. No difference was seen (p>0.05) between the mean prevalence rate of T. gondii of herds to which cats had access as compared to herds that had no contact with cats using a Ttest. However, the mean prevalence rate in farms where cats had access to the herd was 31.01% as compared to 17.34% in farms where there were no cats. Access of dogs was observed in 100% of the farms...

Anticorpos IgG anti-neospora caninum e toxoplasma gondii em búfalas (Bubalus bubalis) criadas no estado do Pará / Occurrence of IgG antibodies anti-neospora caninum and toxoplasma gondii in female water buffaloes (Bubalus bubalis) raised in the Brazilian state of Pará

Para determinar a ocorrência de anticorpos IgG anti-Neospora caninum e Toxoplasma gondii em 14 unidades produtivas de búfalos, situadas em 13 municípios no estado do Pará, foram coletadas amostras de soro sanguíneo de 374 fêmeas adultas. Os soros foram submetidos à reação de imunofluorescência indireta (RIFI), utilizando-se os títulos 200 e 64, respectivamente como ponto de corte para N. caninum e T. gondii. Obteve-se 153 (40,9 por cento) de animais soropositivos para N. caninum com 100 por cento das propriedades com focos da infecção, enquanto que quatro búfalas (1,1 por cento) foram soropositivas em quatro fazendas para T. gondii e uma búfala (0,27 por cento) foi soropositiva para ambos parasitos. A presença de anticorpos é um indicativo da circulação desses protozoários em búfalos das propriedades estudadas, representando uma fonte de infecção para outros animais, assim como o possível envolvimento em distúrbios reprodutivos nessa espécie.

To investigate the occurrence of antibodies IgG anti-Neospora caninum and Toxoplasma gondii, blood samples were collected from 374 adult female water buffaloes originated from 14 production units located in 13 counties of the state of Pará. The sera were subjected to indirect fluorescent antibody (IFA), using titers of 200 and 64 as IFA cut off points for N. caninum and T. gondii, respectively. Hundred and fifty three (40.9 percent) buffaloes were sero-positive for N. caninum, with 100 percent of the properties being sources of infection. Four buffaloes (1.1 percent) in four farms were seropositive for T. gondii, and one buffalo cow (0.27 percent) was seropositive for the two parasites. The presence of antibodies indicates that these protozoan parasites are circulating among buffaloes from the properties studied, representing a source of infection to other animals, as well as a possible cause of reproductive disorders in this species.

Occurrence of antibodies to Neospora caninum and Toxoplasma gondii in dairy cattle from the northern region of the Paraná State, Brazil

Foram analisadas, por meio da imunofluorescência indireta, 385 amostras de soros de vacas, pertencentes a 90 propriedades leiteiras de 12 municípios da região Norte do estado do Paraná. Foram observados 45 (12%) sororeagentes ao Neospora caninum e 102 (26%) ao Toxoplasma gondii. Apenas quatro animais apresentaram títulos de anticorpos para ambos os coccídios. Não foi observada diferença significativa na associação entre a sorologia do N. caninum e as variáveis relacionadas ao manejo, produção de leite, problemas reprodutivos, alimentação, presença de cães, gatos e roedores. Os resultados sugerem que neosporose e toxoplasmose estão disseminadas nos rebanhos leiteiros da região Norte do estado do Paraná, e a freqüência simultânea de anticorpos anti-N. caninum e anti-T. gondii, demonstra sua ocorrência independente em vacas leiteiras.

Comparison of three techniques for the serological diagnosis of Neospora caninum in the dog and their use for epidemiological studies.

An enzyme-linked immunosorbent assay (ELISA) was developed in our laboratory and used to determine the seroprevalence of Neospora caninum in three different dog populations in Belgium: healthy dogs from cattle farms and urban dogs with or without various neurological disorders. The test was validated and compared with two other tests: an indirect fluorescent antibody test (IFAT) and a commercially available competitive enzyme-linked immunosorbent assay (C-ELISA). The study showed a good correlation between the IFAT and the ELISA developed. When the two tests were compared with the C-ELISA, moderate positive and negative agreement indices were observed. Using our ELISA and the IFAT techniques, a high prevalence was found in farm dogs. This result showed that the neurological symptoms are not usually associated with the Neospora infection. In conclusion, the ELISA developed in our laboratory could replace the IFAT for the screening of a large number of dogs' sera.

A role for balance of interferon-gamma and interleukin-4 production in protective immunity against Neospora caninum infection.

A suitable balance in the production of Th1/Th2-type cytokines has a crucial role in the control of microbial infections. We investigated cytokine production patterns and effects during Neospora caninum infection, based on two mouse models and an in vitro system. In the acute infection of N. caninum, BALB/c-background IFN-gamma-deficient mice that were sensitive to the N. caninum infection showed high levels of IL-10 production, whereas significant levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production were observed in resistant wild type mice. BALB/c mice vaccinated with recombinant vaccinia virus expressing N. caninum surface protein NcSRS2 resisted parasite spread throughout the body, low levels of IFN-gamma production and high levels of IL-4 production were observed compared to unvaccinated animals. The treatment of N. caninum-infected cells with IFN-gamma or IL-10 decreased the host-cell viability in an in vitro system using mouse macrophage J774A.1 cells. On the other hand, IL-4, but not IL-10 administration, increased the viability of N. caninum-infected and IFN-gamma-treated cells. In the light of the balance of Th1/Th2-type cytokine production, an IFN-gamma/IL-4 balance may have a crucial role for the control of cellular responses against the parasite invasion.

Neospora caninum: is it really different from Hammondia heydorni or is it a strain of Toxoplasma gondii? An opinion.

The published data concerning Toxoplasma gondii, Hammondia hammondi, H. heydorni and Neospora caninum on one side and between T. gondii on the other were neglected by most authors. As conclusion we are convinced that there are only two valid species: Isospora (Toxoplasma) gondii and Hammondia heydorni. The first includes as a strain H. hammondi and the latter N. caninum. In any case there is absolutely no reason (with respect to general Zoological nomenclature) to create new genera!