AVALIAÇÃO MORFOLÓGICA DO SÊMEN DE TATU-CANASTRA (Priodontes maximus Kerr, 1792)

Resumo

O tatu-canastra (Priodontes maximus), classificado como vulnerável à extinção, é uma das espécies mais raras e emblemáticas da fauna sul-americana  ADDIN ZOTERO_ITEM CSL_CITATION
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world’s most comprehensive information source on the global conservation status
of animal, fungi and plant
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(1). A coleta e a criopreservação de sêmen têm sido adotadas como estratégias relevantes em programas de conservação de espécies ameaçadas. No entanto, o conhecimento limitado sobre a fisiologia reprodutiva de P. maximus representa um entrave ao desenvolvimento e à aplicação deste tipo de iniciativas para a conservação desta espécie  ADDIN ZOTERO_ITEM CSL_CITATION
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viable populations of all wildlife species requires the maintenance of habitat,
as well as an understanding of the behaviour and physiology of individual
species. Despite substantial efforts, there are thousands of species threatened
by extinction, often because of complex factors related to politics, social and
environmental conditions and economic needs. When species become critically
endangered, ex situ recovery programmes that include reproductive scientists
are the usual first line of defence. Despite the potential of reproductive
technologies for rapidly increasing numbers in such small populations, there
are few examples of success. This is not the result of a failure on the part of
the technologies per se, but rather is due to a lack of knowledge about the
fundamental biology of the species in question, information essential for
allowing reproductive technologies to be effective in the production of
offspring. In addition, modern conservation concepts correctly emphasise the
importance of maintaining heterozygosity to sustain genetic vigour, thereby
limiting the practical usefulness of some procedures (such as nuclear
transfer). However, because of the goal of maintaining all extant gene
diversity and because, inevitably, many species are (or will become)
'critically endangered', it is necessary to explore every avenue for a
potential contributory role. There are many 'emerging technologies' emanating
from the study of livestock and laboratory animals. We predict that a subset of
these may have application to the rescue of valuable genes from individual
endangered species and eventually to the genetic management of entire
populations or species. The present paper reviews the potential candidate
techniques and their potential value (and limitations) to the study and
conservation of rare wildlife
species.","container-title":"Reproduction, Fertility, and
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of emerging technologies to the study and conservation of threatened and
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(2). Este estudo teve como objetivo avaliar a morfologia espermática de P. maximus de vida-livre. Em outubro de 2025, foram realizadas duas capturas de dois machos adultos, monitorados por GPS e radiotelemetria no Pantanal do Mato Grosso do Sul. Os procedimentos foram autorizados pelo SISBIO (nº 27587-1) e pela CEUA (nº 1919/2023). A anestesia foi induzida por via intramuscular com tartarato de butorfanol, cloridrato de detomidina e midazolam (0,1 mg/kg cada), seguida de cetamina (5 mg/kg)  ADDIN ZOTERO_ITEM CSL_CITATION
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anatomy and physiology of armadillos make anesthetic procedures and the
placement of telemetry devices challenging. From June 2011 to November 2019, a
total of 73 free-living armadillos were captured and anesthetized in the Pantanal,
Brazil. Giant (Priodontes maximus), six-banded (Euphractus sexcinctus),
southern naked-tailed (Cabassous unicinctus), and nine-banded (Dasypus
novemcinctus) armadillos were divided into two groups. In group 1, 30
armadillos were anesthetized for collection of biological samples, body
measurements, and placement of a microchip tag. Anesthetic combination BDM was
applied: butorphanol tartrate, detomidine hydrochloride, and midazolam
hydrochloride, each at 0.1 mg/kg. In group 2, 43 armadillos received ketamine
hydrochloride at 10 mg/kg 20 min after BDM injection, and intra-abdominal radio
transmitters were surgically implanted. The transmitter was inserted freely
into the abdominal cavity. Vital signs were monitored during anesthesia every
10 min and varied within species and between groups. Rectal temperature varied
from 33.1 ± 1.36 to 35.34 ± 1.21°C, heart rate (beats/min) from 19 ± 2.14 to
84.71 ± 9.25, respiratory rate (breaths/min) from 11 ± 4.16 to 31 ± 2.82, and
oxygen saturation values (SPO2%) from 84.17% ± 2.39 to 98% ± 1.20. Both groups
received the antagonist combination NYF: naloxone hydrochloride (0.02 mg/kg),
yohimbine hydrochloride (0.125 mg/kg), and flumazenil (0.01 mg/kg). Recovery
varied according to intravenous or intramuscular injection from 2 ± 4 to 8.08 ±
2.93 min respectively. BDM protocol was considered satisfactory and provided
enough time to complete the procedures (60 ± 85 to 133.20 ± 9.12 min) according
to the species and group. Ketamine added to the BDM provided enough time and a
surgical plane of anesthesia (97 ± 22 to 137 ± 39.5 min). The surgical
procedure technique chosen did not appear to have a negative impact on
armadillos studied. Implantable transmitters provide a cost-effective method
for long-term monitoring of wild individuals.","container-title":"Journal
of Zoo and Wildlife Medicine: Official Publication of the American Association
of Zoo
Veterinarians","DOI":"10.1638/2017-0194","issue":"3","journalAbbreviation":"J
Zoo Wildl
Med","language":"eng","note":"PMID:
33480527","page":"514-526","source":"PubMed","title":"Anesthesia
and surgery protocols for intra-abdominal transmitter placement in four species
of wild armadillo","volume":"51","author":[{"family":"Kluyber","given":"Danilo"},{"family":"Pinho
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Jean
Desbiez","given":"Arnaud"}],"issued":{"date-parts":[["2020",11]]}}}],"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}
(3). A coleta de sêmen foi realizada por eletroejaculação, utilizando uma sonda retal de 17 cm de comprimento e 1,3 cm de diâmetro, contendo dois eletrodos de 7 cm. O protocolo incluiu a aplicação de 80 estímulos elétricos distribuídos em três séries progressivas de voltagem  ADDIN ZOTERO_ITEM CSL_CITATION
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giant armadillo (Priodontes maximus) is the largest living armadillo and is
classified as Vulnerable by the International Union for Conservation of Nature.
Little is known about the species biology and especially its reproduction.
However, data such as the age at sexual maturity are employed for estimation of
a species generation length, which is an important parameter for extinction
risk assessment. Thus, the aim of this study is to estimate the age at sexual
maturity for male P. maximus in the wild and to predict generation length for
this species. During a long-term in situ study, eight male P. maximus were
captured for reproductive evaluation considering both morphometric
characteristics (e.g. body, penis and testes dimensions), and data on sperm
production (assessed by electroejaculation). Based on observations of this
study, it is estimated that male P. maximus reach sexual maturity at an age of
6.5–8 years. This is at least 5.5–7 years later than previously published.
Consequently, generation length for P. maximus results in 11 years when
applying standard formula including age at sexual maturity in males and an
estimate of age at last reproduction of 20 years. However, the generation
length is probably underestimated here as sexual maturity in females and age at
last reproduction could be later than estimates used. Even these conservative
estimates result in noticeable change for the timeframe used in extinction risk
assessments for this species as it is 33 instead of 21 years. Thus, results
from this study might affect the classification status of P. maximus
particularly for regional
assessments.","container-title":"Mammalian Biology -
Zeitschrift fur Saugetierkunde","DOI":"10.1007/s42991-020-00065-3","issue":"932","journalAbbreviation":"Mammalian
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Saugetierkunde","language":"en","source":"ResearchGate","title":"Size
matters: penis size, sexual maturity and their consequences for giant armadillo
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(4). O sêmen foi colhido em tubos de 2 mL e fixado em solução formol-salina tamponada, sendo posteriormente armazenado a 4 °C até à realização das análises morfológicas.
A avaliação morfológica dos espermatozoides foi realizada por microscopia óptica (objetiva de 1000x), após a coloração de esfregaços de sêmen fixado pelo método Karras modificado  ADDIN ZOTERO_ITEM CSL_CITATION
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armadillos (Euphractus sexcinctus), a non-endangered species, serve as a
valuable experimental model for developing assisted reproductive technologies
aimed at conserving endangered species within the order Cingulata. However,
limited knowledge of their reproductive biology presents a critical challenge.
This study aimed to evaluate the effectiveness of a pharmacological semen
collection protocol and compare Karras-modified staining using brightfield
microscopy (KA), phase contrast microscopy (PC), and differential interference
contrast microscopy (DIC) for assessing sperm morphology. Twenty free-ranging
male six-banded armadillos were captured and subjected to the following
pharmacological sedation protocol: butorphanol tartrate, detomidine
hydrochloride and midazolam hydrochloride (i.m., 0.1 mg/kg each). Ten minutes
after administration, semen was extracted by applying gentle pressure along the
penis, from its base to the tip. The expelled semen, which accumulated into the
urethral fossa, was collected using a variable-volume micropipette and assessed
for macro and microscopic characteristics. Ninety-five percent of the males
exhibited a partial ejaculation, with no aversive behavior or signs of
discomfort observed. Most semen samples (83 %) contained more than 30 % of
morphologically abnormal sperm with considerable individual variability in
sperm defect profiles. While all microscopy techniques allowed effective
assessment of sperm morphology, KA was the most efficient in identifying
acrosome integrity and head abnormalities. This method offers a practical and
cost-effective solution, particularly suited for field conditions. This study
reports the first successful pharmacological protocol for semen collection in
six-banded armadillos, highlighting its potential use for reproductive
assessments and contributing to developing conservation strategies for related
species.","container-title":"Animal Reproduction
Science","DOI":"10.1016/j.anireprosci.2025.107851","journalAbbreviation":"Animal
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Science","language":"en","page":"107851","source":"ScienceDirect","title":"Pharmacologically
assisted semen collection and sperm morphology assessment methods in wild
six-banded armadillos (Euphractus
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Carlos
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Grego"},{"family":"Desbiez","given":"Arnaud
L. J."},{"family":"De
Camilo","given":"Beatriz
Lippe"},{"family":"Da
Silva","given":"Luan
Sitó"},{"family":"De
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(5). Para cada amostra, foram analisadas 200 células quanto à morfologia e 100 quanto à morfometria, considerando-se o comprimento e a largura da cabeça e o comprimento da cauda. Os espermatozoides apresentaram cabeças volumosas e arredondadas, com o acrossoma em forma de ferradura, semelhante ao padrão descrito para o tatu-peba (Euphractus sexcinctus)  ADDIN ZOTERO_ITEM CSL_CITATION
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armadillos (Euphractus sexcinctus), a non-endangered species, serve as a
valuable experimental model for developing assisted reproductive technologies
aimed at conserving endangered species within the order Cingulata. However,
limited knowledge of their reproductive biology presents a critical challenge.
This study aimed to evaluate the effectiveness of a pharmacological semen
collection protocol and compare Karras-modified staining using brightfield
microscopy (KA), phase contrast microscopy (PC), and differential interference
contrast microscopy (DIC) for assessing sperm morphology. Twenty free-ranging
male six-banded armadillos were captured and subjected to the following
pharmacological sedation protocol: butorphanol tartrate, detomidine
hydrochloride and midazolam hydrochloride (i.m., 0.1 mg/kg each). Ten minutes
after administration, semen was extracted by applying gentle pressure along the
penis, from its base to the tip. The expelled semen, which accumulated into the
urethral fossa, was collected using a variable-volume micropipette and assessed
for macro and microscopic characteristics. Ninety-five percent of the males
exhibited a partial ejaculation, with no aversive behavior or signs of
discomfort observed. Most semen samples (83 %) contained more than 30 % of
morphologically abnormal sperm with considerable individual variability in
sperm defect profiles. While all microscopy techniques allowed effective
assessment of sperm morphology, KA was the most efficient in identifying
acrosome integrity and head abnormalities. This method offers a practical and
cost-effective solution, particularly suited for field conditions. This study
reports the first successful pharmacological protocol for semen collection in
six-banded armadillos, highlighting its potential use for reproductive
assessments and contributing to developing conservation strategies for related
species.","container-title":"Animal Reproduction
Science","DOI":"10.1016/j.anireprosci.2025.107851","journalAbbreviation":"Animal
Reproduction
Science","language":"en","page":"107851","source":"ScienceDirect","title":"Pharmacologically
assisted semen collection and sperm morphology assessment methods in wild
six-banded armadillos (Euphractus
sexcinctus)","volume":"277","author":[{"family":"Lobo","given":"Ana
Carolina
M."},{"family":"Ferreira","given":"João
Carlos
Pinheiro"},{"family":"Kluyber","given":"Danilo"},{"family":"Caiaffa","given":"Mayara
Grego"},{"family":"Desbiez","given":"Arnaud
L. J."},{"family":"De
Camilo","given":"Beatriz
Lippe"},{"family":"Da
Silva","given":"Luan
Sitó"},{"family":"De
Camargo","given":"Gabriel
C."},{"family":"Kasimanickam","given":"Ramanathan"},{"family":"Oba","given":"Eunice"}],"issued":{"date-parts":[["2025",6,1]]}}}],"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}
(5). Foram observados agrupamentos celulares do tipo “rouleaux”, compostos por duas a três células (Figura 1C). Os defeitos mais frequentes incluíram cabeças isoladas normais (Figura 1D), caudas enroladas ou dobradas (Figura 1A) e gotas citoplasmáticas distais (Figura 1B), que demonstraram elevada afinidade pela técnica de coloração utilizada. As médias morfométricas (± SEM) foram: comprimento da cabeça - 19,48 ± 0,41 µm; largura da cabeça - 16,87 ± 0,36 µm; e comprimento da cauda - 75,47 ± 1,86 µm. Os resultados indicam que o método Karras modificado foi eficiente para a caracterização estrutural dos espermatozoides de P. maximus, embora a identificação do acrossoma tenha se mostrado mais desafiadora em comparação com E. sexcinctus  ADDIN ZOTERO_ITEM CSL_CITATION
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armadillos (Euphractus sexcinctus), a non-endangered species, serve as a
valuable experimental model for developing assisted reproductive technologies
aimed at conserving endangered species within the order Cingulata. However,
limited knowledge of their reproductive biology presents a critical challenge.
This study aimed to evaluate the effectiveness of a pharmacological semen
collection protocol and compare Karras-modified staining using brightfield
microscopy (KA), phase contrast microscopy (PC), and differential interference
contrast microscopy (DIC) for assessing sperm morphology. Twenty free-ranging
male six-banded armadillos were captured and subjected to the following
pharmacological sedation protocol: butorphanol tartrate, detomidine
hydrochloride and midazolam hydrochloride (i.m., 0.1 mg/kg each). Ten minutes
after administration, semen was extracted by applying gentle pressure along the
penis, from its base to the tip. The expelled semen, which accumulated into the
urethral fossa, was collected using a variable-volume micropipette and assessed
for macro and microscopic characteristics. Ninety-five percent of the males
exhibited a partial ejaculation, with no aversive behavior or signs of
discomfort observed. Most semen samples (83 %) contained more than 30 % of
morphologically abnormal sperm with considerable individual variability in
sperm defect profiles. While all microscopy techniques allowed effective
assessment of sperm morphology, KA was the most efficient in identifying
acrosome integrity and head abnormalities. This method offers a practical and
cost-effective solution, particularly suited for field conditions. This study
reports the first successful pharmacological protocol for semen collection in
six-banded armadillos, highlighting its potential use for reproductive
assessments and contributing to developing conservation strategies for related
species.","container-title":"Animal Reproduction
Science","DOI":"10.1016/j.anireprosci.2025.107851","journalAbbreviation":"Animal
Reproduction
Science","language":"en","page":"107851","source":"ScienceDirect","title":"Pharmacologically
assisted semen collection and sperm morphology assessment methods in wild
six-banded armadillos (Euphractus
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Carlos
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Grego"},{"family":"Desbiez","given":"Arnaud
L. J."},{"family":"De
Camilo","given":"Beatriz
Lippe"},{"family":"Da
Silva","given":"Luan
Sitó"},{"family":"De
Camargo","given":"Gabriel
C."},{"family":"Kasimanickam","given":"Ramanathan"},{"family":"Oba","given":"Eunice"}],"issued":{"date-parts":[["2025",6,1]]}}}],"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}
(5). Além disso, apesar das dimensões superiores dos componentes celulares, a morfologia espermática de P. maximus apresenta uma notável semelhança com a de E. sexcinctus  ADDIN ZOTERO_ITEM CSL_CITATION
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armadillos (Euphractus sexcinctus), a non-endangered species, serve as a
valuable experimental model for developing assisted reproductive technologies
aimed at conserving endangered species within the order Cingulata. However,
limited knowledge of their reproductive biology presents a critical challenge.
This study aimed to evaluate the effectiveness of a pharmacological semen
collection protocol and compare Karras-modified staining using brightfield
microscopy (KA), phase contrast microscopy (PC), and differential interference
contrast microscopy (DIC) for assessing sperm morphology. Twenty free-ranging
male six-banded armadillos were captured and subjected to the following
pharmacological sedation protocol: butorphanol tartrate, detomidine
hydrochloride and midazolam hydrochloride (i.m., 0.1 mg/kg each). Ten minutes
after administration, semen was extracted by applying gentle pressure along the
penis, from its base to the tip. The expelled semen, which accumulated into the
urethral fossa, was collected using a variable-volume micropipette and assessed
for macro and microscopic characteristics. Ninety-five percent of the males
exhibited a partial ejaculation, with no aversive behavior or signs of
discomfort observed. Most semen samples (83 %) contained more than 30 % of
morphologically abnormal sperm with considerable individual variability in
sperm defect profiles. While all microscopy techniques allowed effective
assessment of sperm morphology, KA was the most efficient in identifying
acrosome integrity and head abnormalities. This method offers a practical and
cost-effective solution, particularly suited for field conditions. This study
reports the first successful pharmacological protocol for semen collection in
six-banded armadillos, highlighting its potential use for reproductive
assessments and contributing to developing conservation strategies for related
species.","container-title":"Animal Reproduction
Science","DOI":"10.1016/j.anireprosci.2025.107851","journalAbbreviation":"Animal
Reproduction
Science","language":"en","page":"107851","source":"ScienceDirect","title":"Pharmacologically
assisted semen collection and sperm morphology assessment methods in wild
six-banded armadillos (Euphractus
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Carlos
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Grego"},{"family":"Desbiez","given":"Arnaud
L. J."},{"family":"De
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Sitó"},{"family":"De
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(5), sugerindo o potencial desta espécie como modelo experimental para o desenvolvimento de protocolos de preservação de sêmen de P. maximus. Os achados morfológicos e morfométricos dos espermatozóides descritos neste estudo contribuem, de forma inédita, para o avanço do conhecimento sobre a reprodução do tatu-canastra, fornecendo subsídios científicos relevantes para ações de conservação da espécie.