Clinical and radiological results after one hundred fifteen MPFL reconstructions with or without tibial tubercle transfer in patients with recurrent patellar dislocation-a mean follow-up of 5.4 years.

PURPOSE: Medial patellofemoral ligament reconstruction (MPFLR) shows low recurrent rates and high satisfaction. Reports on outcome at mid-term follow-up in a large cohort with or without tibial tubercle transfer (TTT) are scarce. METHODS: One hundred six patients (115 knees; 24.3 ± 8.7 years) with recurrent patellar dislocation underwent MPFLR with ipsilateral gracilis autograft and were included in this retrospective follow-up study. In 43 knees, simultaneous TTT was performed for patellar maltracking due to patella alta or increased tibial tubercle trochlear groove distance (TT-TG). Kujala and subjective knee scores were recorded pre- and post-operatively. RESULTS: Eighty-seven percent of the patients were satisfied at a mean follow-up of 5.4 ± 3.1 years. The Kujala score increased in 81.7% of all knees from 50.9 to 76.2 points (p < 0.01). Patients with a decreased Kujala score were not different in terms of follow-up time, age, trochlear dysplasia, or post-operative patellar height. The decline was related to an increase in pain, whereas functional scores remained identical. There were six patients with persisting instability, three with patellar re-dislocation. Re-operation was necessary in 24 patients (21.2%), mainly for implant removal after TTT and loss of flexion. Performing TTT had no significant influence on the subjective outcome. Patients with more than eight years of follow-up (n = 27) did not show any difference in the subjective outcome parameters, or in osteoarthritis progression. CONCLUSION: MPFLR with and without TTT is a reliable treatment option for recurrent patellar dislocation without deterioration at mid-term follow-up. Persisting pain is a major post-operative issue that seems to be unrelated to patellofemoral anatomy.

[Bizarre parosteal osteochondromatous proliferation of the metatarsal bone]. / Bizarre paraosseale osteochondromatöse Proliferation der Metatarsalknochen.

A bizarre parosteal osteochondromatous proliferation (BPOP), also known as a Nora lesion, is a rare benign bone formation. It emanates mainly from the intact cortex of the metaphysis of short tubular bones (hands or feet). Conventional radiographs should be complemented using cross-sectional imaging modalities (CT/MRI). In the absence of symptoms a non-operative regime with radiological and clinical controls is possible. If symptomatic, excision biopsy is the treatment of choice, though a high recurrence rate has been reported. Histopathological examination confirms the diagnosis and typically shows an endochondral ossification zone and an unusually mineralized cartilaginous matrix referred to as "blue bone".

Factors affecting reproductive success in three entomophilous orchid species in Hungary.

The reproductive success of orchids is traditionally estimated by determining the fruit-set of individuals. Here, we investigated both the fruit and the seed production of three orchid species and the factors that may affect individual fruit-set, like pollination strategy, individual traits or the annual amount of precipitation. The species [Dactylorhiza sambucina (L.) Soó, Dactylorhiza majalis (Rchb.) P. F. Hunt & Summerhayes and Platanthera bifolia (L.) L. C. M. Richard] were studied in three consecutive years (2010-2012) in the Bükk Mountains, Hungary. All three species were proved to be non-autogamous by a bagging experiment. Data analyses showed significant differences between seed numbers but not between fruit-sets of species. There was no statistical difference in individual reproductive success between wet and dry years, however, the effect of the annual amount of precipitation is significant on the population level. Comparison of published fruit-set data revealed accordance with our results in P. bifolia, but not in D. sambucina and D. majalis. We assume that the surprisingly high fruit-set values of the two Dactylorhiza species may be due to the fact that the pollination crisis reported from Western European countries is not an actual problem in the Bükk Mountains, Hungary.

Molecular phylogeny and evolutionary history of the Eurasiatic orchid genus Himantoglossum s.l. (Orchidaceae).

BACKGROUND AND AIMS: Lizard orchids of the genus Himantoglossum include many of Eurasia's most spectacular orchids, producing substantial spikes of showy flowers. However, until recently the genus had received only limited, and entirely traditional, systematic study. The aim of the current work was to provide a more robust molecular phylogeny in order to better understand the evolutionary relationships among species of particular conservation concern. METHODS: All putative species of Himantoglossum s.l. were sampled across its geographical range. A large subsample of the 153 populations studied contributed to an initial survey of nuclear ribosomal internal transcribed spacer (nrITS) ribotypes. Smaller subsets were then sequenced for four plastid regions and the first intron of the low-copy-number nuclear gene LEAFY. Rooted using Steveniella as outgroup, phylogenetic trees were generated using parsimony and Bayesian methods from each of the three datasets, supplemented with a ribotype network. KEY RESULTS: The resulting trees collectively determined the order of branching of the early divergent taxa as Himantoglossum comperianum > H. robertianum group > H. formosum, events that also involved significant morphological divergence. Relaxed molecular clock dating suggested that these divergences preceded the Pleistocene glaciations (the origin of the H. robertianum group may have coincided with the Messinian salinity crisis) and occurred in Asia Minor and/or the Caucasus. Among more controversial taxa of the H. hircinum-jankae clade, which are only subtly morphologically divergent, topological resolution was poorer and topological incongruence between datasets was consequently greater. CONCLUSIONS: Plastid sequence divergence is broadly consistent with prior, morphologically circumscribed taxa and indicates a division between H. hircinum-adriaticum to the west of the Carpathians and H. jankae-caprinum (plus local endemics) to the east, a distinction also suggested by nrITS ribotypes. LEAFY phylogenies are less congruent with prior taxonomic arrangements and include one likely example of paralogy. Himantoglossum metlesicsianum fully merits its IUCN Endangered status. Potentially significant genetic variation was detected within Steveniella satyrioides, H. robertianum and H. hircinum. However, confident circumscription of the more derived species of Himantoglossum s.s., including local endemics of hybrid origin, must await future morphometric and population-genetic analyses.

Copper Intoxication in South American Camelids-Review of the Literature and First Report of a Case in a Vicuña (Vicugna vicugna).

Copper (Cu), an essential trace element in the metabolism of mammals, plays a central role in various metabolic processes. However, overdosing can lead to severe symptoms and even fatalities. Chronic Cu intoxication continues to be a problem in grazing and domestic animals, with sheep being particularly sensitive. There are few comparative studies on its impact on South American camelids (SACs). Therefore, this work presents the results of literature research combined with a case report on a 3-year-old female vicuña (Vicugna vicugna) presented to the clinic from a zoological garden in northern Germany. The animal showed reduced food intake, recumbency, bruxism, icteric mucous membranes and sclera. Auscultation revealed atony of the third compartment and the digestive tract. Similar to cases described in the literature, the animal showed rapid deterioration of its condition with unspecific symptoms of liver failure and rapid death. However, in contrast to descriptions in sheep, clinical icterus has not been previously reported in cases of other SACs. Laboratory findings from EDTA and serum samples revealed neutrophilia with a left shift, hypoproteinaemia, lymphopaenia, azotaemia, elevated levels of creatine kinase (CK), aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH) in the serum. Hyperbilirubinaemia and significantly elevated serum and liver Cu levels were observed. Subsequent blood samples from the remaining vicuñas and alpacas in the same enclosure showed no remarkable abnormalities. To the best of the authors' knowledge, this case report represents the first documented case of Cu intoxication specifically in vicuñas.

The draft genome of Spiraea crenata L. (Rosaceae) - the first complete genome in tribe Spiraeeae.

Spiraea crenata L. is a deciduous shrub distributed across the Eurasian steppe zone. The species is of cultural and horticultural importance and occurs in scattered populations throughout its westernmost range. Currently, there is no genomic information on the tribe of Spiraeeae. Therefore we sequenced and assembled the whole genome of S. crenata using second- and third-generation sequencing and a hybrid assembly approach to expand genomic resources for conservation and support research on this horticulturally important lineage. In addition to the organellar genomes (the plastome and the mitochondrion), we present the first draft genome of the species with an estimated size of 220 Mbp, an N50 value of 7.7 Mbp, and a BUSCO score of 96.0%. Being the first complete genome in tribe Spiraeeae, this may not only be the first step in the genomic study of a rare plant but also a contribution to genomic resources supporting the study of biodiversity and evolutionary history of Rosaceae.

New thousand-seed weight dataset for plant species of Central Europe.

One of the most important and most easily measurable physical characteristics of plant seeds is their weight, which influences and indicates crucial ecological processes. Seed weight affects spatial and temporal dispersibility, and can also influence seed predation and the germination, growth and survival of seedlings. Providing trait data for species missing from international databases is key to promote studies that advance our understanding of the functioning of plant communities and ecosystems, which is an essential issue in the face of the global climate change and biodiversity loss. Compared to species from Western and Northwestern Europe, those with an Eastern or Central European centre of distribution are underrepresented in most international trait databases. Therefore, the creation of specific trait databases is key to help regional studies. In this respect, it is important not only to collect fresh seeds for weight measurements, but also to measure and process data of seeds preserved in collections and make them available to the broader scientific community. In this data paper we provide seed weight data to fill in missing trait data of plant species of Central and Eastern Europe. Our dataset includes weight measurement for 281 taxa of the Central European flora including also some cultivated and exotic species. The seeds were collected between 1971 and 2021 mostly in Central Europe. One part of the measured seeds was collected in the last decade, the other part is from an older seed collection, but all seeds were measured recently. For each species, we collected a minimum of 3 × 100 intact seeds, if possible. The seeds were air-dried at room temperature (approximately 21 °C and 50% relative humidity) for at least two weeks and measured with an accuracy of 0.001 g using an analytical balance. The thousand-seed weights reported here were calculated based on the measured values. Our goal for the future is to incorporate the seed weight data reported here in a regional database (Pannonian Database of Plant Traits - PADAPT) that gathers plant traits and other plant characteristics for the Pannonian flora. The data presented here will facilitate trait-based analyses of the flora and vegetation of Central Europe.

PADAPT 1.0 - the Pannonian Dataset of Plant Traits.

The existing plant trait databases' applicability is limited for studies dealing with the flora and vegetation of the eastern and central part of Europe and for large-scale comparisons across regions, mostly because their geographical data coverage is limited and they incorporate records from several different sources, often from regions with markedly different climatic conditions. These problems motivated the compilation of a regional dataset for the flora of the Pannonian region (Eastern Central Europe). PADAPT, the Pannonian Dataset of Plant Traits relies on regional data sources and collates data on 54 traits and attributes of the plant species of the Pannonian region. The current version covers approximately 90% of the species of the region and consists of 126,337 records on 2745 taxa. By including species of the eastern part of Europe not covered by other databases, PADAPT can facilitate studying the flora and vegetation of the eastern part of the continent. Although data coverage is far from complete, PADAPT meets the longstanding need for a regional database of the Pannonian flora.

Road traffic and landscape characteristics predict the occurrence of native halophytes on roadside verges.

Road management practices, such as winter de-icing create ideal habitats and competitive advantage for salt-tolerant species. We aimed to map the occurrences of halophytes along roads in Hungary. Furthermore, we tested factors that might play a role in the roadside occurrences of five chosen native halophytes from rare to common, we encountered during our field surveys. These were Festuca pseudovina, Limonium gmelinii subsp. hungaricum, Podospermum canum, Puccinellia distans and Spergularia media. We found, that at least one halophyte species was documented in 71% of the total sampling points. Germination experiments indicated that substrate salt concentration significantly decreased germination rates in each of the five species, but in case of L. gmelinii subsp. hungaricum, or P. distans germination occurred on extremely high salt concentrations. Traffic intensity, the presence of other halophytes at the sampling point and the presence of a given species in the surrounding landscape had a significant positive effect on the occurrence of four of the five model species. Our results suggest that the studied species are mostly in the early stage of their roadside spread, colonizing roadsides close to their native distribution ranges. The possibility of a future range expansion along roads cannot be excluded.

Trade of commercial potting substrates: A largely overlooked means of the long-distance dispersal of plants.

Although long-distance dispersal (LDD) events are rare and stochastic, they are disproportionately important and drive several large-scale ecological processes; yet, we have a very limited understanding of their frequency, extent and consequences. Humanity intentionally spreads several species, which is associated with the accidental dispersal of other plant species. Although the global trade of potted plants and horticultural substrates may disperse large quantities of propagules, it has hardly been studied from an ecological point of view. We assessed the viable seed content of different types of commercial potting substrates to answer the following questions: (i) In what richness and density do substrates contain viable seeds? (ii) Does the composition of substrates influence their viable seed content? and (iii) Are there common characteristics of the species dispersed this way? We detected 438 seedlings of 66 taxa and found that 1 l of potting substrate contains an average of 13.27 seeds of 6.24 species, so an average 20-liter bag of substrate contains 265 viable seeds. There was a high variability in the seed content of the substrates, as substrates containing manure contained a substantially higher number of species and seeds than substrates without manure. Thus, this pathway of LDD is an interplay between endozoochory by grazing livestock and accidental human-vectored dispersal, implying that the diet preference of grazing animals influences the ability of a plant species to be dispersed this way. According to our results, potting substrates can disperse large quantities of seeds of a wide range of plant species over large distances. We conclude that this kind of human-vectored LDD may have complex effects on plant populations and communities; however, as this dispersal pathway is largely understudied and has hardly been considered as a type of LDD, its consequences are still unknown and further studies of the issue are of great importance.

Population pharmacokinetic analysis of ropivacaine and its metabolite 2',6'-pipecoloxylidide from pooled data in neonates, infants, and children.

BACKGROUND: The aim was to characterize ropivacaine and 2',6'-pipecoloxylidide (PPX) pharmacokinetics and factors affecting them in paediatric anaesthesia. METHODS: Population pharmacokinetics of ropivacaine and its active metabolite PPX were estimated after single and continuous ropivacaine blocks in 192 patients aged 0-12 yr from six pooled published studies. Unbound and total ropivacaine and PPX plasma concentration and PPX urinary excretion data were used for non-linear mixed-effects modelling by NONMEM. Covariates included age, body weight, gender, ethnic origin, ASA, site and method of administration, and total dose. RESULTS: One-compartment first-order pharmacokinetic models incorporating linear binding of ropivacaine and PPX to α(1)-acid glycoprotein were used. After accounting for the effect of body weight, clearance of unbound ropivacaine and PPX reached 41% and 89% of their mature values, respectively, at the age of 6 months. Ropivacaine half-life decreased with age from 13 h in the newborn to 3 h beyond 1 yr. PPX half-life differed from 19 h in the newborn to 8-11 h between 1 and 12 months to 17 h after 1 yr. Simulations indicate that for a single caudal block, the recommended dose could be increased by a factor of 2.9 (0-1 month group) and 6.3 (1-12 yr group) before the unbound plasma concentrations would cross the threshold for systemic toxicity. Corresponding factors for continuous epidural infusion are 1.8 and 4.9. CONCLUSIONS: Ropivacaine and PPX unbound clearance depends on body weight and age. The results support approved dose recommendations of ropivacaine for the paediatric population.

Diterpene alkaloids from Aconitum anthora and assessment of the hERG-inhibiting ability of Aconitum alkaloids.

A new norditerpene alkaloid, 10-hydroxy-8- O-methyltalatizamine (1), was isolated from the whole plant of ACONITUM ANTHORA L. besides the known isotalatizidine (2) and hetisinone (3). The structures were determined by means of HR-ESI-MS, 1D and 2D NMR spectroscopy, including ¹H-¹H COSY, NOESY, HSQC and HMBC experiments, resulting in complete ¹H and ¹³C chemical shift assignments for 1- 3, and revision of some earlier ¹³C-NMR data. The effects of the isolated compounds, together with twenty-one other ACONITUM alkaloids with different skeletal types and substitution patterns, on hERG channels were studied by the whole-cell patch clamp technique, using the QPatch-16 automated patch clamp system. At 10 µM, aconitine, 14-benzoylaconine 8- O-palmitate, songoramine, gigactonine and neolinine demonstrated significant hERG K+ channel inhibition; all other compounds exerted only low (6-21%) inhibitory activity.

Orchids of Azerbaijani Cemeteries.

In order to explore their orchid flora, we performed surveys of 96 Azerbaijani burial places in 2018 and 2019. Altogether, 28 orchid taxa were found in 37 visited cemeteries. In the orchid diversity a remarkable pattern was observed: geographic latitude was significantly and positively related to the number of taxa and number of individuals. The most widespread and abundant orchids in Azerbaijani graveyards were Anacamptis pyramidalis and A. papilionacea (found in 23 and 8 cemeteries, respectively). Azerbaijani cemeteries can be important refuges for rare and threatened orchids, e.g., Himantoglossum formosum (three cemeteries), Ophrys sphegodes subsp. mammosa (eight), Orchis adenocheila (two), O. punctulata (three), O. stevenii (one) and Steveniella satyrioides (one). Epipactis turcica, detected in a single locality, was previously unknown to the flora of Azerbaijan. Additionally, we documented orchid tuber (salep) collection in two cemeteries.

[Is postoperative acute pain control in colorectal surgery better within an enhanced recovery after surgery program (ERAS)?] / ¿Mejora el control del dolor postoperatorio en cirugía colorrectal tras la implementación de un protocolo de rehabilitación intensificada (ERAS)?

BACKGROUND AND OBJECTIVE: A good acute pain control is necessary to achieve the main objective of Enhanced recovery After Surgery (ERAS) programs and accelerate recovery. The study objective is to evaluate postoperative (PO) acute pain, PO opioid consumption, and its association with functional recovery, after implementing a colorectal surgery ERAS program. An analysis was made as regards drugs adverse effects. METHODS: Observational cohort study on scheduled colorectal Surgery: one prospective cohort subjected to the ERAS care program, and one retrospective cohort that received traditional non-standardised care. A record was made of mean pain intensity (measured by a visual analogue scale, which classifies pain intensity from 0 to 10, from lower to higher intensity), as well as the amount of opioid consumption on the day of surgery and on the first three postoperative days, and drugs adverse effects. An analysis was made of the association between PO opioid consumption and ERAS program, and between PO opioid consumption and functional recovery. RESULTS: The study included a total of 410 patients (313 in the ERAS group and 97 in the control group). In the ERAS group, it was observed that the mean visual analogue scale was less than 2, with a smaller amount of PO opioid consumption, on each single day and the accumulated amount of the four days (4 [0-24] vs. 0 [0-4], P<.001). PO opioid consumption was associated with functional recovery (OR 0.97 [95% CI; 0.96-0.99], P=.011). No drugs adverse effects were observed. CONCLUSIONS: After implementing a colorectal Surgery ERAS program, good pain control was achieved, as well as a reduction in PO opioid consumption, which is associated with functional recovery. No drugs adverse effects were observed.

Entamoeba ranarum Infection in a Ball Python (Python regius).

The pathogenic Entamoeba species in snakes is widely regarded to be Entamoeba invadens, which can cause severe amoebiasis with up to 100% mortality. In this case report, we describe a ball python (Python regius) that died after short-term weight loss. Necropsy revealed severe necrotizing colitis with large numbers of intralesional Entamoeba trophozoites. Molecular genetic analysis identified these trophozoites as Entamoeba ranarum, a parasite more usually found in amphibians. Furthermore, the extended history revealed that toads (Rhinella marina) had been housed together with the python. This report illustrates the danger of protozoal cross-infections in exotic animals as well as the importance of molecular genetic tools in Entamoeba diagnosis.

Occurrence of the rare plant Sternbergia colchiciflora in an urban environment.

In this paper, we report a remarkable population of the rare plant Sternbergia colchiciflora found along a busy road section in the downtown of county seat Veszprém (W-Hungary). The population contains at least 7000 individuals and spreads across fifteen grassy traffic islands. Regarding the position of individuals, their relative distance from the road/sidewalk within the traffic islands differed significantly from a hypothetical uniform distribution, with higher number of individuals situated close to the island edges than expected by chance. Besides this rare bulbous plant, several other dry grassland specialist plant species were also found in these habitat patches, e.g., Astragalus austriacus, Festuca rupicola, Filipendula vulgaris, Muscari neglectum, Petrorhagia saxifraga, Potentilla arenaria, Ranunculus illyricus, Salvia nemorosa, Sanguisorba minor, Teucrium chamaedrys, Thesium linophyllon, Verbascum phoeniceum and Vinca herbacea. The origin of these dry grassland plants in this highly disturbed urban environment is uncertain; possibly, they were introduced ca. 15 years ago in the course of earthworks during the construction of the road. Recent management (frequent and motorized lawn mowing) seems to favor the Sternbergia colchiciflora, while the habitat is currently not directly threatened. Unfortunately, the potential for local conservation appears to be highly limited.

Roadsides provide refuge for orchids: characteristic of the surrounding landscape.

Seminatural habitats are declining throughout the world; thus, the role of small anthropogenic habitats in the preservation of plants is becoming increasingly appreciated. Here, we surveyed the orchid flora of roadside verges in five Central European countries (Austria, Hungary, Romania, Slovakia, and Slovenia) and tested how the surrounding landscape matrix affects the overall number of species and individuals, and also different functional groups of orchids. We found more than 2,000 individuals of 27 orchid species during our surveys. According to our results, the increasing coverage of agricultural and urban areas negatively affects both the number of orchid species and individuals on roadsides. Our study further suggests that differences in the surrounding habitats affect which species are found on roadsides, since the increasing coverage of grasslands or forested areas around orchid occurrences had a significant positive effect on the number of grassland or forest-dwelling species and individuals, respectively. Most variance in orchid numerosity and diversity was explained by the cover of the suitable habitat types of the respective taxa in the surrounding landscape of the sampling points. This highlights the importance of roadsides acting as refugia for numerous species and valuable plant communities as well as in supporting biodiversity in general.

The protected flora of long-established cemeteries in Hungary: Using historical maps in biodiversity conservation.

The role of anthropogenically influenced habitats in conserving elements of the original wildlife has increased worldwide simultaneously with the disappearance of natural sites. Burial places are able to conserve original elements of the wildlife, and this fact has been known for at least a century. To this day, little is known about long-time changes and the effect of long-time management methods in cemeteries on the flora they harbor. The utility of historical maps in research focused on natural values, as well as in answering questions related to conservation was recently demonstrated, but the use of digitized historical maps in biodiversity research of the Carpathian Basin is very limited. In the present paper, we aimed to predict the conservation potential of long-established and newly established cemeteries of Hungarian settlements with various population sizes based on the digitized maps of the 2nd Military Survey of the Austrian Empire (1819-1869), by categorizing cemeteries into 3 distinct (anthropogenic habitat, cemetery, or natural habitat) types. To build our models, we used records of the protected flora from Hungarian cemeteries, based on data of thematic botanical surveys of 991 cemeteries. Out of the surveyed cemeteries, 553 (56%) harbored protected plants, totaling 306.617 estimated individuals of 92 protected species, belonging to 28 plant families. These species represent 12% of the entire protected flora of Hungary. Hungarian cemeteries play a key role mainly in preserving steppe and dry grassland plant species. Long-established and large cemeteries harbor more protected plant species than small and newly established ones. Human population size of the settlements correlated negatively with the number of protected species and individuals. Moreover, woodland cover and proportion of grassland also significantly positively affected the number of protected plant species in cemeteries.

Seed mass, hardness, and phylogeny explain the potential for endozoochory by granivorous waterbirds.

Field studies have shown that waterbirds, especially members of the Anatidae family, are major vectors of dispersal by endozoochory for a broad range of plants lacking a fleshy fruit, yet whose propagules can survive gut passage. Widely adopted dispersal syndromes ignore this dispersal mechanism, and we currently have little understanding of what traits determine the potential of angiosperms for endozoochory by waterbirds. Results from previous experimental studies have been inconsistent as to how seed traits affect seed survival and retention time in the gut and have failed to control for the influence of plant phylogeny. Using 13 angiosperm species from aquatic and terrestrial habitats representing nine families, we examined the effects of seed size, shape, and hardness on the proportion of seeds surviving gut passage through mallards (Anas platyrhynchos) and their retention time within the gut. We compiled a molecular phylogeny for these species and controlled for the nonindependence of taxa due to common descent in our analyses. Intact seeds from all 13 species were egested, but seed survival was strongly determined by phylogeny and by partial effects of seed mass and hardness (wet load): species with seeds harder than expected from their size, and smaller than expected from their loading, had greater survival. Once phylogeny was controlled for, a positive partial effect of seed roundness on seed survival was also revealed. Species with seeds harder than expected from their size had a longer mean retention time, a result retained after controlling for phylogeny. Our study is the first to demonstrate that seed shape and phylogeny are important predictors of seed survival in the avian gut. Our results demonstrate that the importance of controlling simultaneously for multiple traits and relating single traits (e.g., seed size) alone to seed survival or retention time is not a reliable way to detect important patterns, especially when phylogenetic effects are ignored.

First successful artificial insemination with frozen-thawed semen in rhinoceros.

The first successful artificial insemination (AI) in a rhinoceros was reported in 2007 using fresh semen. Following that success, we decided to evaluate the possibility of using frozen-thawed semen for artificial insemination. Semen, collected from a 35-36 year old Southern white rhinoceros (Ceratotherium simum simum) in the UK was frozen using the directional freezing technique. This frozen semen was used in two intrauterine AI attempts on a 30 years old female rhinoceros in Hungary. The first attempt, conducted 30 days postpartum with an insemination dose of approximately 135 x 10(6) motile cells, failed. The second attempt, conducted two estrus cycles later with an insemination dose of approximately 500 x 10(6) motile cells, resulted in pregnancy and the birth of a healthy offspring. This represents the first successful AI using frozen-thawed semen in a rhinoceros, putting it among very few wildlife species in which AI with frozen-thawed semen resulted in a live birth. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or between wild and captive populations, without the need to transport stressed or potentially disease carrying animals. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for managing genetic diversity in these endangered mammals.